Beta-lactamase inhibitor compounds

ABSTRACT

The present invention is directed to compounds which are beta-lacatamase inhibitors. The compounds and their pharmaceutically acceptable salts, are useful in combination with beta-lactam antibiotics, or alone, for the treatment of bacterial infections, including infections caused by drug resistant organisms, including multi-drug resistant organisms. The present invention includes compounds according to formula (Ia): 
                         
or a pharmaceutically acceptable salt thereof, wherein the values of R 1 , R 2 , R 3  and R 4  are described herein.

FIELD OF THE INVENTION

The present invention relates to novel beta-lactamase inhibitors, theirpharmaceutical compositions and methods of use. In addition, the presentinvention relates to therapeutic methods for the treatment of bacterialinfections, including overcoming bacterial antibiotic resistance.

BACKGROUND OF THE INVENTION

The international microbiological and infectious disease communitycontinues to express serious concern that the continuing evolution ofantibacterial resistance could result in bacterial strains against whichcurrently available antibacterial agents will be ineffective. Theoutcome of such an occurrence could have considerable morbidity andmortality. In general, bacterial pathogens may be classified as eitherGram-positive or Gram-negative pathogens. Antibiotic compounds witheffective activity against both Gram-positive and Gram-negativepathogens are typically regarded as having a broad spectrum of activity.

In the fight against bacterial infection, beta-lactam antibiotics areessential. Beta-lactams are a broad class of drugs which all have abeta-lactam in their core molecular structure, and typically showeffectiveness against a broad spectrum of Gram-positive andGram-negative bacteria by inhibiting the cell wall synthesis of thebacterium. Because the drug target has no eukaryotic analog, theirtoxicity is low and they are generally well-tolerated. They remain amongthe most widely prescribed, safe and effective drugs available to combatbacterial infection. However, their effectiveness is limited by highlyresistant infectious strains such as methicillin-resistantStaphylococcus aureus (MRSA) and multi-drug resistant (MDR) strains ofPseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli,Klebsiella pneumonia, and other Enterobacteriaceae. Such resistantbacteria are major causes of patient morbidity and mortality. Helfand,β-lactams Against Emerging ‘Superbugs’: Progress and Pitfalls, ExpertRev. Clin. Pharmacol. 1(4):559-571 (2008).

Beta-lactam antibiotics, alone and in combination with beta-lactamaseinhibitors, continue to represent an essential portion of theantibacterial agents used to combat disease. β-lactam resistance forGram-negative infections is primarily driven by β-lactamase activity;and the significant dependence on β-lacatam antibiotics has lead to thediversification and increased prevalence of β-lactamases. Theseβ-lactamases are driving resistance to even the newest β-lactamantibiotics. Llarrull, et al., The Future of Beta-Lactams, CurrentOpinion in Microbiology, 13:551-557 (2010).

A major threat to the efficacy of these drugs is the increasingprevalence of extended-spectrum beta-lactamases (ESBLs). Beta-lactamasesare enzymes that are secreted by some bacteria that ring open thebeta-lactam portion of a beta-lactam antibiotic and thereby deactivateit. There are currently, four classes of beta-lactamases, denoted ClassA, Class B, Class C and Class D. Class A, Class C and Class Dbeta-lactamases are serine beta-lactamase inhibitors, while Class Bbeta-lactamases are metallo-beta-lactamases (MBLs). Bush & Jacoby,Updated Functional Classification of β-Lactamases, Antimicrobial Agentsand Chemotherapy, 54(3):969-976 (March 2010).

To help improve the effectiveness of beta-lactam antibiotics, somebeta-lactamase inhibitors have been developed. However, the currentlyavailable β-lactamase inhibitors in many instances are insufficient tocounter the constantly increasing diversity of β-lactamases. The threemost common serine beta-lactamase agents currently used—clavulanic acid,tazobactam and sulbactam—have activity only against certain Class Aenzymes, which severely limits their utility. Additionally,beta-lactamase inhibitors currently in clinical trials, such asAvibactam and MK7655 work primarily on Class A and C enzymes, withminimal effectiveness against Class D beta-lactamases. Bebrone, et al.,Current Challenges in Antimicrobial Chemotherapy: Focus on β-LactamaseInhibition, Drugs, 70(6):651-679 (2010). While these agents represent aconsiderable improvement over the currently available beta-lactamaseinhibitors, agents which effectively hit all three serinebeta-lactamases are desirable for combating the significant beta-lactamresistance seen today. Currently, there are no approved β-lactamaseinhibitors which are effective against Class D β-lactamases, andresistance rates to conventional antibiotics are continuing to rise.

Therefore, there is a need for new β-lactamase inhibitors which areeffective against at least D β-lactamases. There is a clear need for newβ-lactamase inhibitors which are effective against more than one ofClass A, C and/or D β-lactamases.

SUMMARY OF THE INVENTION

The present invention is directed to compounds which are beta-lacatamaseinhibitors. The compounds, and their pharmaceutically acceptable salts,are useful in combination with beta-lactam antibiotics, or alone, forthe treatment of bacterial infections, including infections caused bydrug resistant organisms, including multi-drug resistant organisms. Moreparticularly, the invention relates to compounds of formula (Ia):

or a pharmaceutically acceptable salt thereof, wherein R¹ is —CONR′R″,—CN, or C₁-C₃ alkyl, wherein each alkyl is optionally substituted withC₁-C₃ alkoxy, —OH, —CN, —NR′R″, or —CONR′R″; R² and R³ are independentlyselected from H, halo, —CN, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₆ cycloalkyl, C₁-C₆ alkoxy, —CONR′R″, or C(O)₂R′; wherein the alkyl,alkenyl, cycloalkyl, and alkoxy represented by R² or R³ areindependently and optionally substituted by one or more halo, —CN, —OH,C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃haloalkoxy, —NR′R″, 5-7 membered heterocycle, —C(O)NR′R″ or —NR′C(O)R″;and each R′ and R″ are independently selected from hydrogen, C₁-C₆alkyl, C₃-C₆ cycloalkyl, phenyl, 5 to 6 membered heterocyclyl or a 5 to6 membered heteroaryl; wherein each alkyl, cycloalkyl, phenyl,heterocyclyl and heteroaryl is optionally and independently substitutedwith one or more halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₃-C₆cycloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —C(O)(C₁-C₆ alkyl),—C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃ alkyl), —N(C₁-C₃ alkyl)₂, a 5-7membered heterocyclyl or a 5-7 membered heteroaryl; provided that R² andR³ are not both hydrogen; and when R¹ is —C(O)NR′R″, then neither of R²or R³ is —C(O)NR′R″.

DETAILED DESCRIPTION OF THE INVENTION

In one aspect of the invention is a beta-lactamase inhibitor compoundaccording to formula (I):

or a pharmaceutically acceptable salt thereof, wherein R¹ is —CONR′R″,—CN, C₁-C₃ alkyl or C₁-C₂ alkoxy, wherein each alkyl and alkoxy isindependently and optionally substituted with C₁-C₃ alkoxy, —OH, —CN,—NR′R″, —CONR′R″ or a 5-7 membered heterocycle; R² and R³ areindependently selected from H, halo, —CN, C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₁-C₆ alkoxy, —CONR′R″, C(O)₂R′,phenyl, 5-6 membered heterocyclyl or 5-6 membered heteroaryl; whereinthe alkyl, alkenyl, cycloalkyl, alkoxy, phenyl, heterocyclyl andheteroaryl represented by R² or R³ are independently and optionallysubstituted by one or more halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl,C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″, 5-7 memberedheterocyclyl, —C(O)NR′R″ or —NR′C(O)R″; R⁴ is —OS(O)₂OH, —S(O)₂OH,—OP(O)₂OH, —P(O)₂OH, —C(O)NHS(O)₂R⁵, —OCHFCO₂H, —OCF₂CO₂H, or —OCH₂CO₂H;R⁵ is NR′R″, phenyl, a 5-6 membered heterocyclyl or a 5 to 6 memberedheteroaryl; and each R′ and R″ is independently selected from hydrogen,C₁-C₆ alkyl, C₃-C₆ cycloalkyl, phenyl, 5-7 membered heterocyclyl or a5-6 membered heteroaryl; wherein each alkyl, cycloalkyl, phenyl,heterocyclyl and heteroaryl is optionally and independently substitutedwith one or more halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₃-C₆cycloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —C(O)(C₁-C₆ alkyl),—C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃ alkyl), —N(C₁-C₃ alkyl)₂, a 5-6membered heterocyclyl or a 5-6 membered heteroaryl; or R′ and R″ aretaken together to form a 5-6 membered heterocyclyl or heteroaryl,wherein each heterocyclyl and heteroaryl is optionally and independentlysubstituted with one or more halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NH₂,—NH(C₁-C₃ alkyl), or —N(C₁-C₃ alkyl)₂; provided that R² and R³ are notboth hydrogen; and when R¹ is —C(O)NR′R″, then neither R² nor R³ is—C(O)NR′R″.

In another aspect of the invention is a compound according to formula(II):

or a pharmaceutically acceptable salt thereof wherein the variables R¹,R², R³ and R⁴ are as defined for formula (I) above.

In one aspect of the invention is a compound according to formula (III):

or a pharmaceutically acceptable salt thereof, wherein the variables R¹,R² and R³ are as defined for formula (Ia) above.

In one aspect of the invention, is a compound according to formula (IV):

or a pharmaceutically acceptable salt thereof, wherein the variables R¹and R³ are as defined for formula (Ia).

One aspect of the invention is a compound according to formula (V):

or a pharmaceutically acceptable salt thereof, wherein the variables R¹and R² are as defined for formula (Ia).In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), R¹ is —CONR′R″, —CN, or C₁-C₃ alkyl, wherein eachalkyl is optionally substituted with C₁-C₃ alkoxy or —OH; and the R′ andR″ of R¹ are independently selected from the group consisting of H,C₁-C₃ alkyl, or a 5-7 membered heterocyclyl, wherein each alkyl andheterocyclyl of R′ and R″ is optionally and independently substitutedwith one or more —OH, C₁-C₃ alkyl, C₁-C₃ alkoxy, —NH₂, —NH(C₁-C₃ alkyl),—N(C₁-C₃ alkyl)₂, or a 5-7 membered heterocyclyl. In one aspect of theinvention, for any one of formulae (I), (Ia), (II), (III), (IV) or (V),R¹ is —CH₂OCH₃, —CONH(CH₂)-siderophore, —CONH₂,

and

represents the point of attachment to the bridged bicyclic core. In oneaspect of the invention, for any one of formulae (I), (Ia), (II), (III),(IV) or (V), R¹ is —CH₂OCH₃ or —CONH₂.

In one aspect of the invention, for any of formulae (I), (II), (III), or(IV), R¹ is —C(O)NH₂, —CN or C₁-C₃ alkyl optionally substituted with oneor more —OH, C₁-C₃ alkoxy, halo, —OC(O)NR′R″, a siderophore, or—C(O)NH(siderophore), wherein R′ and R″ are as defined for any one offormulae (I), (Ia), (II), (III), (IV) or (V). In one aspect of theinvention, for any one of formulae (I), (II), (III) or (IV), R¹ is—C(O)NH₂, —CN or C₁-C₂ alkyl optionally substituted with methoxy, —OH or—CN. In one aspect of the invention, for any one of formulae (I), (Ia),(II), (III), (IV) or (V), R¹ is —CONR′R″, —CH₂OCH₃ or —CN; and the R′and R″ of R¹ are independently —H, C₁-C₃ alkyl, or a 5-7 memberedheterocyclyl, wherein each alkyl and heterocyclyl of R′ and R″ isoptionally and independently substituted with one or more halo, —CN,—OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,—C(O)(C₁-C₆ alkyl), —C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃ alkyl),—N(C₁-C₃ alkyl)₂, or a 5-7 membered heterocyclyl. In one aspect of theinvention, for any one of formulae (I), (Ia), (II), (III), (IV) or (V),R¹ is —CONH₂, —CONH(CH₂)_(n)NH₂,

n is an integer from 1 to 3; and

represents the point of attachment to the bridged bicyclic core. In oneaspect of the invention, for any one of formulae (I), (Ia), (II), (III),(IV) or (V), R¹ is —CONH₂,

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), R¹ is —CONH₂. In one aspect of the invention, forany one of formulae (I), (Ia), (II), (III), (IV) or (V), R¹ is —CH₂OCH₃.In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), R¹ is —CN. In one aspect of the invention, for anyone of formulae (I), (Ia), (II), (III), (IV) or (V), R¹ is —CH₂OH.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III) or (V), R² is selected from the group consisting of H, C₁-C₃alkyl, C₃-C₆ cycloalkyl, and —CONR′R″, wherein the alkyl and cycloalkylrepresented by R² and/or R³ are independently and optionally substitutedby one or more group selected from halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″, a siderophore,—C(O)NR′R″ and —NR′C(O)R″. In one aspect of the invention, for any oneof formulae (I), (Ia), (II), (III) or (V), R² is methyl, ethyl,isopropyl, or cyclopropyl, wherein each R² is optionally andindependently substituted with one or more group selected from —OH andC₁-C₃ alkoxy. In one aspect of the invention, for any one of formulae(I), (Ia), (II), (III) or (V), R² is methyl.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III) or (V), R² is —H, —CN, C₁-C₃ alkyl, C₃-C₆ cycloalkyl, —CO₂R′,—CONR′R″, or a 5-6 membered heterocyclyl, wherein each alkyl,cycloalkyl, heterocyclyl, R′ and R″ of R² is optionally andindependently substituted with one or more group selected from halo,—CN, —OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,—NR′R″, morpholinyl, pyrrolidinyl, piperidinyl, piperazinyl, asiderophore, —C(O)NR′R″ and —NR′C(O)R″. In one aspect of the invention,for any one of formulae (I), (Ia), (II), (III) or (V), R² is H, —CN,methyl, ethyl, isopropyl, cyclopropyl, —CO₂(C₁-C₃ alkyl), —CONH₂,—CONH(C₁-C₃ alkyl), —CON(C₁-C₃ alkyl)₂, morpholinyl or thiazolyl,wherein when R² is not hydrogen or cyano, each R² is optionally andindependently substituted with one or more group selected from halo,—CN, —OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,—NR′R″, morpholinyl, pyrrolidinyl, piperidinyl, piperazinyl, asiderophore, —C(O)NR′R″ and —NR′C(O)R″. In one aspect of the invention,for any one of formulae (I), (Ia), (II), (III) or (V), R² is H, —CN,methyl, ethyl, propyl, isopropyl, thiazolyl, —CONR′R″, or —CO₂CH₃,wherein when R² is not hydrogen or cyano, each R² is optionally andindependently substituted by one or more fluoro, chloro, bromo, C₁-C₃alkyl, C₁-C₃ haloalkyl, C₁-C₃ alkoxy or —NR′R″; and R′ and R″, whenpresent in R², are independently selected from H and methyl. In oneaspect of the invention, for any one of formulae (I), (Ia), (II), (III)or (V), R² is H, —CN, methyl, isopropyl, —CONHCH₃, —CONH(CH₂)₂NH₂,—CO₂CH₃, —(CH₂)NH₂, —(CH₂)₂NH₂ or thiazolyl. In one aspect of theinvention, for any one of formulae (I), (Ia), (II), or (III), R² ishydrogen.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III) or (IV), R³ is selected from the group consisting of H, C₁-C₃alkyl, C₃-C₆ cycloalkyl, and —CONR′R″, wherein the alkyl and cycloalkylrepresented by R² and/or R³ are independently and optionally substitutedby one or more group selected from halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″, a siderophore,—C(O)NR′R″ and —NR′C(O)R″. In one aspect of the invention, for any oneof formulae (I), (Ia), (II), (III) or (IV), R³ is C₁-C₃ alkyl, C₂-C₆alkenyl, C₃-C₆ cycloalkyl, or —CONR′R″, each of which is optionally andindependently substituted with one or more substituent selected from thegroup consisting of halo, —CN, —OH, C₁-C₃ alkyl, cyclopropyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″, a siderophore,—C(O)NR′R″ and —NR′C(O)R″; and each R′ and R″ is independently selectedfrom H and C₁-C₃ alkyl. In one aspect of the invention, for any one offormulae (I), (Ia), (II), (III) or (IV), R³ is methyl, ethyl, isopropyl,cyclopropyl, —CONH₂, —CONH(C₁-C₃ alkyl), or —CON(C₁-C₃ alkyl)₂, each ofwhich is optionally and independently substituted with one or more groupselected from —OH, C₁-C₃ alkyl, C₁-C₃ alkoxy, —NR′R″, C(O)NR′R″ and—NR′C(O)R″; and each R′ and R″ is independently selected from H andC₁-C₃ alkyl. In one aspect of the invention, for any one of formulae(I), (Ia), (II), (III) or (IV), R³ is C₁-C₃ alkyl, cyclopropyl,—CONR′R″, wherein each alkyl, and cyclopropyl is optionally andindependently substituted with one or more —OH, C₁-C₃ alkoxy, —NH₂, or—NHC(O)(C₁-C₃ alkyl); and each R′ and R″ are independently selected fromH, C₁-C₃ alkyl, and 5-6 membered heterocyclyl, wherein each alkyl andheterocyclyl represented by R′ or R″ is optionally and independentlysubstituted with one or more —OH, C₁-C₃ alkyl, or C₁-C₃ alkoxy. In oneaspect of the invention, for any one of formulae (I), (Ia), (II), (III)or (IV), R³ is methyl, —CH₂OCH₃, or —CONH₂.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III) or (IV), R³ is H, C₁-C₃ alkyl, C₃-C₆ cycloalkyl, —CO₂R′, —CONR′R″,or a 5-6 membered heterocyclyl, each of which is optionally andindependently substituted with one or more group selected from halo,—CN, —OH, C₁-C₃ alkyl, cyclopropyl, C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃haloalkoxy, —NR′R″, morpholinyl, pyrrolidinyl, piperidinyl, piperazinyl,—C(O)NR′R″ and —NR′C(O)R″; and wherein each R′ and R″, when present inR³, is independently selected from H and C₁-C₃ alkyl. In one aspect ofthe invention, for any one of formulae (I), (Ia), (II), (III) or (IV),R³ is H, methyl, ethyl, isopropyl, cyclopropyl, —CO₂(C₁-C₃ alkyl),—CONH₂, —CONH(C₁-C₃ alkyl), —CON(C₁-C₃ alkyl)₂, morpholinyl orthiazolyl, each of which is optionally and independently substitutedwith one or more group selected from halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃haloalkyl, cyclopropyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″,morpholinyl, pyrrolidinyl, piperidinyl, piperazinyl, —C(O)NR′R″ and—NR′C(O)R″; and wherein, when present in R³, each R′ and R″ isindependently selected from H and C₁-C₃ alkyl. In one aspect of theinvention, for any one of formulae (I), (Ia), (II), (III) or (IV), R³ isH, C₁-C₃ alkyl, C₂-C₄ alkenyl, C₃-C₆ cycloalkyl, —CONR′R″, or aheterocyclyl, wherein each alkyl, alkenyl, and heterocyclyl isoptionally and independently substituted with one or more halo, —CN,—OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,—C(O)(C₁-C₆ alkyl), —C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃ alkyl), or—N(C₁-C₃ alkyl)₂; and when present in R³, each R′ and R″ are optionallyand independently substituted with one or more halo, —CN, —OH, C₁-C₃alkyl, C₁-C₃ haloalkyl, cyclopropyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,—C(O)(C₁-C₆ alkyl), —C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃ alkyl), or—N(C₁-C₃ alkyl)₂. In one aspect of the invention, for any one offormulae (I), (Ia), (II), (III) or (IV), R³ is —CONR′R″; and one of R′and R″ is H and the other is C₁-C₃ alkyl optionally substituted with oneor more halo, —CN, —OH, —CF₃, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,cyclopropyl, —C(O)(C₁-C₆ alkyl), —C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃alkyl), or —N(C₁-C₃ alkyl)₂. In one aspect of the invention, for any oneof formulae (I), (Ia), (II), (III) or (IV), R³ is —CONH(CH₂)_(n)NHR′; R′is H, methyl, ethyl, propyl, isopropyl or cyclopropyl; and n is aninteger from 1-3. In one aspect of the invention, for any one offormulae (I), (Ia), (II), (III) or (IV), R³ is —CONH(CH₂)₂NH₂. In oneaspect of the invention, for any one of formulae (I), (Ia), (II), (III)or (IV), R³ is methyl, isopropyl, isopropenyl, —CONH₂ or —CON(CH₃)₂. Inone aspect of the invention, for any one of formulae (I), (Ia), (II),(III) or (IV), R³ is —CH₂OCH₃. In one aspect of the invention, for anyone of formulae (I), (Ia), (II), or (III), R³ is hydrogen.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), each R′ and R″ are independently selected fromhydrogen, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, phenyl, 5 to 6 memberedheterocyclyl or a 5 to 6 membered heteroaryl; wherein each alkyl,cycloalkyl, phenyl, heterocyclyl and heteroaryl is optionally andindependently substituted with one or more halo, —CN, —OH, C₁-C₃ alkyl,C₁-C₃ haloalkyl, C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,—C(O)(C₁-C₆ alkyl), —C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃ alkyl),—N(C₁-C₃ alkyl)₂, a 5-7 membered heterocyclyl or a 5-7 memberedheteroaryl. In one aspect of the invention, for any one of formulae (I),(Ia), (II), (III), (IV) or (V), each R′ and R″ are independentlyselected from H, C₁-C₃ alkyl, and 5-6 membered heterocyclyl, whereineach alkyl and heterocyclyl represented by R′ or R″ is optionally andindependently substituted with one or more —OH, C₁-C₃ alkyl, or C₁-C₃alkoxy. In one aspect of the invention, for any one of formulae (I),(Ia), (II), (III), (IV) or (V), each R′ and R″ is independently selectedfrom H and C₁-C₃ alkyl.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), each R′ and R″ is independently selected fromhydrogen, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, phenyl, 5-7 memberedheterocyclyl or a 5-6 membered heteroaryl; wherein each alkyl,cycloalkyl, phenyl, heterocyclyl and heteroaryl is optionally andindependently substituted with one or more halo, —CN, —OH, C₁-C₃ alkyl,C₁-C₃ haloalkyl, C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy,—C(O)(C₁-C₆ alkyl), —C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃ alkyl),—N(C₁-C₃ alkyl)₂, a 5-6 membered heterocyclyl or a 5-6 memberedheteroaryl; or R′ and R″ are taken together to form a 5-6 memberedheterocyclyl or heteroaryl, wherein each heterocyclyl and heteroaryl isoptionally and independently substituted with one or more halo, —CN,—OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃haloalkoxy, —NH₂, —NH(C₁-C₃ alkyl), or —N(C₁-C₃ alkyl)₂. In one aspectof the invention, for any one of formulae (I), (Ia), (II), (III), (IV)or (V), each R′ and R″ is independently selected from hydrogen, andC₁-C₆ alkyl. In one aspect of the invention, for any one of formulae(I), (Ia), (II), (III), (IV) or (V), each R′ and R″ is independentlyselected from hydrogen, methyl, ethyl, propyl and isopropyl. In oneaspect of the invention, for any one of formulae (I), (Ia), (II), (III),(IV) or (V), each R′ and R″ is independently selected from hydrogen andmethyl. In one aspect of the invention, for any one of formulae (I),(Ia), (II), (III), (IV) or (V), each R′ and R″ is independently selectedfrom an C₁-C₃ alkyl optionally substituted with one or more of methoxy,ethoxy, —OH, —NH₂, NH(CH₃), —N(CH₃)₂, or a siderophore. In one aspect ofthe invention, for any one of formulae (I), (Ia), (II), (III), (IV) or(V), one of R′ and R″ is hydrogen, while the other is selected from anyof the possible values listed above.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), R¹ is —CH₂OCH₃; —CONH₂, or

R² is —H or —CH₃; and R³ is —H, —CH₃, or —CONH₂; provided that R² and R³are not both H; and when R¹ is —CONH₂, or

then R³ is not —CONH₂.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), R¹ is —CH₂OCH₃; —CONH₂,

R² is —H or methyl; R³ is —H, —CH₃, or —CONH₂; and R⁴ is —OSO₂OH.

In one aspect of the invention, for any one of formulae (I), (Ia), (II),(III), (IV) or (V), R¹ is —CH₂OCH₃; —CONR′R″,

R² is —H, C₁-C₃ alkyl or C₃-C₆ cycloalkyl; R³ is —H, C₁-C₃ alkyl, C₃-C₆cycloalkyl or —CONR′R″; R⁴ is —OSO₂OH; and each R′ and R″ areindependently —H or C₁-C₃ alkyl.

In either of the two above aspects of the invention, the compound is asdefined, provided that R² and R³ are not both H; and when R¹ is —CONH₂,

then R³ is not —CONH₂.

One aspect of the invention is the compound:

or a pharmaceutically acceptable salt thereof.

One aspect of the invention is the compound:

or a pharmaceutically acceptable salt thereof.

One aspect of the invention is the compound:

or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention is the compound:

or a pharmaceutically acceptable salt thereof.

One aspect of the invention is the compound:

or a pharmaceutically acceptable salt thereof.

One aspect of the invention is the compound:

or a pharmaceutically acceptable salt thereof.

Any embodiment described herein can be combined with any other suitableembodiment described herein to provide additional embodiments. Forexample, where one embodiment individually or collectively describespossible groups for R¹ and a separate embodiment describes possiblegroups for R², it is understood that these embodiments can be combinedto provide an additional embodiment utilizing the possible groups for R¹with the possible groups for R². Analogously, the invention encompassesany embodiments called out individually for R¹, R², R³, R⁴, R⁵, R′ andR″ in combination with any specific embodiments called out for each ofthe remaining variables.

Compounds of Formulae (I), (Ia), (II), (III), (IV) and (V) possessbeneficial efficacious, metabolic, toxicological and/or pharmacodynamicproperties.

In one aspect of the invention, the compound of formula (Ia) is selectedfrom the group consisting of:

-   (2S,5R)-2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-cyano-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-4-methyl-7-oxo-2-(piperidinium-4-ylcarbamoyl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sulfate;-   (2S,5R)-2-carbamoyl-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-cyano-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-(2-aminoethylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate;-   (2S,5R)-2-(methoxymethyl)-7-oxo-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-((5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-carbamoyl-4-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sodium sulfate;-   (2S,5R)-2-carbamoyl-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sodium sulfate;-   (2S,5R)-4-carbamoyl-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate, monosodium salt;-   (2S,5R)-2,4-bis(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sulfate Sodium salt;-   (2S,5R)-2-(1-(tert-butoxycarbonyl)piperidin-4-ylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sulfate sodium salt;-   (2S,5R)-4-(dimethylcarbamoyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sulfate sodium salt;-   (2S,5R)-2-(hydroxymethyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate Sodium Salt;-   (2S,5R)-3-methyl-7-oxo-2-(piperidin-1-ium-4-ylcarbamoyl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sulfate;-   (2S,5R)-2-carbamoyl-3-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-4-(2-amino-2-oxoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-4-carbamoyl-2-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium Salt;-   (2S,5R)-2-carbamoyl-3,4-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-carbamoyl-3-ethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-4-(2-aminoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate;-   (2S,5R)-2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-4-(2-acetamidoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    sulfate sodium salt;-   (2S,5R)-2-(methoxymethyl)-4-(methylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-2-carbamoyl-4-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;-   (2S,5R)-3-(2-methoxyethyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt; and-   (2S,5R)-2-(((1,5-dihydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl)carbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-yl    hydrogen sulfate sodium salt;    -   or a pharmaceutically acceptable salt thereof.

Alkyl—As used herein the term “alkyl” refers to both straight andbranched chain saturated hydrocarbon radicals having the specifiednumber of carbon atoms. References to individual alkyl groups such as“propyl” are specific for the straight chain version only and referencesto individual branched chain alkyl groups such as ‘isopropyl’ arespecific for the branched chain version only. In one aspect, “alkyl” ismethyl.

Alkenyl—As used herein, the term “alkenyl” refers to both straight andbranched chain hydrocarbon radicals having the specified number ofcarbon atoms and containing at least one carbon-carbon double bond. Forexample, “C₂₋₆alkenyl” includes groups such as C₂₋₅alkenyl, C₂₋₄alkenyl,ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl,5-hexenyl, 2-heptenyl, and 2-methyl-1-heptenyl.

Alkynyl—As used herein, the term “alkynyl” refers to both straight andbranched chain hydrocarbon radicals having the specified number ofcarbon atoms and containing at least one carbon-carbon triple bond. Forexample, “C₂₋₈alkynyl” includes groups such as C₂₋₆alkynyl, C₂₋₄alkynyl,ethynyl, 2-propynyl, 2-methyl-2-propynyl, 3-butynyl, 4-pentynyl,5-hexynyl, 2-heptynyl, and 4-methyl-5-heptynyl.

Halo—As used herein, the term “halo” is intended to include fluoro,chloro, bromo and iodo. In one aspect, the “halo” may refer fluoro,chloro, and bromo. In another aspect, “halo” may refer to fluoro andchloro. In still another aspect, “halo” may refer to fluoro. In yetanother aspect, “halo” may refer to chloro.

Cycloalkyl—In one aspect, “cycloalkyl” refers to a saturated orpartially saturated monocyclic carbon ring, of which one or more —CH₂—groups may be optionally replaced with a corresponding number of —C(O)—groups. Illustrative examples of “cycloalkyl” include cyclopropyl,cyclobutyl, cyclopentyl, and cyclopentenyl. In one aspect, “3- to5-membered carbocyclyl” may be cyclopropyl.

5-7 Membered Heterocyclyl—The term “5-7 membered heterocyclyl” refers toa saturated or partially saturated, non-aromatic monocyclic ringcontaining 5 to 7 ring atoms, of which at least one ring atom isselected from nitrogen, sulfur, and oxygen, and of which a —CH₂— groupmay be optionally replaced by a —C(O)— group. Analogously, “5-6 memberedheterocyclyl” refers to a saturated or partially saturated, non-aromaticmonocyclic ring containing 5 to 6 ring atoms, of which at least one ringatom is selected from nitrogen, sulfur, and oxygen, and of which a —CH₂—group may be optionally replaced by a —C(O)— group. Unless otherwisespecified, “5-7 membered heterocyclyl” and “5-6 membered heterocyclyl”groups may be carbon or nitrogen linked. Ring nitrogen atoms may beoptionally oxidized to form an N-oxide. Ring sulfur atoms may beoptionally oxidized to form S-oxides or sulphones. Illustrative examplesof “5-7 membered heterocyclyl” and “5-6 membered heterocyclyl” include,but are not limited to, azetidinyl, dioxidotetrahydrothiophenyl,2,4-dioxoimidazolidinyl, 3,5-dioxopiperidinyl, furanyl, imidazolyl,isothiazolyl, isoxazolyl, morpholinyl, oxazolyl, oxetanyl,oxoimidazolidinyl, 3-oxo-1-piperazinyl, 2-oxopyrrolidinyl,2-oxotetrahydrofuranyl, oxo-1,3-thiazolidinyl, piperazinyl, piperidyl,2H-pyranyl, pyrazolyl, pyridinyl, pyrrolyl, pyrrolidinyl, pyrimidinyl,pyrazinyl, pyrazolyl, pyridazinyl, 4-pyridonyl, tetrahydrofuranyl,tetrahydropyranyl, thiazolyl, 1,3,4-thiadiazolyl, thiazolidinyl,thiomorpholinyl, thiophenyl, 4H-1,2,4-triazolyl, pyridine-N-oxidyl,tetrazolyl, oxadiazolyl, triazolyl, pyrazinyl, triazinyl, andhomopiperidinyl. In one embodiment, the terms “5-7 memberedheterocyclyl” and “5-6 membered heterocyclyl” includes siderophores of5-7 or 5-6 members which contain at least one heteroatom.

5- or 6-Membered Heteroaryl—The term “5-6 membered heteroaryl” is refersto a monocyclic, aromatic heterocyclyl ring containing 5 or 6 ringatoms, of which at least one ring atom is selected from nitrogen,sulfur, and oxygen. Unless otherwise specified, “5-6 memberedheteroaryl” groups may be carbon or nitrogen linked. Ring nitrogen atomsmay be optionally oxidized to form an N-oxide. Ring sulfur atoms may beoptionally oxidized to form S-oxides. Illustrative examples of “5-6membered heteroaryl” include furanyl, imidazolyl, isothiazolyl,isoxazole, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl,pyridinyl, pyrrolyl, tetrazolyl, thiadiazolyl, thiazolyl, thiophenyl,and triazolyl.

6-Membered Heteroaryl—In one aspect, “heterocyclyl,” 5- or “6-memberedheterocyclyl,” “6-membered heterocyclyl,” and “5- or 6-memberedheteroaryl” may be “6-membered heteroaryl.” The term “6-memberedheteroaryl” is intended to refer to a monocyclic, aromatic heterocyclylring containing 6 ring atoms. Ring nitrogen atoms may be optionallyoxidized to form an N-oxide. Illustrative examples of “6-memberedheteroaryl” include pyrazinyl, pyridazinyl, pyrimidinyl, and pyridinyl.

Siderophore—In one aspect, a “siderophore” is a low molecular weightmoiety that can bind ferric iron. Once bound, these “iron carriers” canfacilitate transport of the molecule into a bacterial cell. The term“siderophore” includes, but is not limited to the followingheterocyclyls:

Optionally substituted—As used herein, the phrase “optionallysubstituted” indicates that substitution is optional and therefore it ispossible for the designated group to be either substituted orunsubstituted. In the event a substitution is desired, the appropriatenumber of hydrogens on the designated group may be replaced with aselection from the indicated substituents, provided that the normalvalency of the atoms on a particular substituent is not exceeded, andthat the substitution results in a stable compound.

In one aspect, when a particular group is designated as being optionallysubstituted with one or more substituents, the particular group may beunsubstituted. In another aspect, the particular group may bear onesubstituent. In another aspect, the particular substituent may bear twosubstituents. In still another aspect, the particular group may bearthree substituents. In yet another aspect, the particular group may bearfour substituents. In a further aspect, the particular group may bearone or two substituents. In still a further aspect, the particular groupmay be unsubstituted, or may bear one or two substituents.

Pharmaceutically Acceptable—As used herein, the phrase “pharmaceuticallyacceptable” refers to those compounds, materials, compositions, and/ordosage forms which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of human beings and animalswithout excessive toxicity, irritation, allergic response, or otherproblem or complication, commensurate with a reasonable benefit/riskratio.

Effective Amount—As used herein, the phrase “effective amount” means anamount of a compound or composition which is sufficient enough tosignificantly and positively modify the symptoms and/or conditions to betreated (e.g., provide a positive clinical response). The effectiveamount of an active ingredient for use in a pharmaceutical compositionwill vary with the particular condition being treated, the severity ofthe condition, the duration of the treatment, the nature of concurrenttherapy, the particular active ingredient(s) being employed, theparticular pharmaceutically-acceptable excipient(s)/carrier(s) utilized,and like factors within the knowledge and expertise of the attendingphysician.

Leaving Group—As used herein, the phrase “leaving group” is intended torefer to groups readily displaceable by a nucleophile such as an aminenucleophile, and alcohol nucleophile, or a thiol nucleophile. Examplesof suitable leaving groups include halo, such as fluoro, chloro, bromo,and sulfonyloxy group, such as methanesulfonyloxy andtoluene-4-sulfonyloxy.

Protecting Group—As used herein, the term “protecting group” is intendedto refer to those groups used to prevent selected reactive groups (suchas carboxy, amino, hydroxy, and mercapto groups) from undergoingundesired reactions. Illustrative examples of suitable protecting groupsfor a hydroxy group include acyl groups; alkanoyl groups such as acetyl;aroyl groups, such as benzoyl; silyl groups, such as trimethylsilyl; andarylmethyl groups, such as benzyl. The deprotection conditions for theabove hydroxy protecting groups will necessarily vary with the choice ofprotecting group. Thus, for example, an acyl group such as an alkanoylor an aroyl group may be removed, for example, by hydrolysis with asuitable base such as an alkali metal hydroxide, for example lithium orsodium hydroxide. Alternatively a silyl group such as trimethylsilyl maybe removed, for example, by fluoride or by aqueous acid; or anarylmethyl group such as a benzyl group may be removed, for example, byhydrogenation in the presence of a catalyst such as palladium-on-carbon.Illustrative examples of suitable protecting groups for an amino groupinclude acyl groups; alkanoyl groups such as acetyl; alkoxycarbonylgroups, such as methoxycarbonyl, ethoxycarbonyl, and t-butoxycarbonyl;arylmethoxycarbonyl groups, such as benzyloxycarbonyl; and aroyl groups,such benzoyl. The deprotection conditions for the above amino protectinggroups necessarily vary with the choice of protecting group. Thus, forexample, an acyl group such as an alkanoyl or alkoxycarbonyl group or anaroyl group may be removed for example, by hydrolysis with a suitablebase such as an alkali metal hydroxide, for example lithium or sodiumhydroxide. Alternatively an acyl group such as a t-butoxycarbonyl groupmay be removed, for example, by treatment with a suitable acid ashydrochloric, sulfuric, phosphoric acid or trifluoroacetic acid and anarylmethoxycarbonyl group such as a benzyloxycarbonyl group may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon, or by treatment with a Lewis acid, for exampleboron trichloride). A suitable alternative protecting group for aprimary amino group is, for example, a phthaloyl group, which may beremoved by treatment with an alkylamine, for exampledimethylaminopropylamine or 2-hydroxyethylamine, or with hydrazine.Another suitable protecting group for an amine is, for example, a cyclicether such as tetrahydrofuran, which may be removed by treatment with asuitable acid such as trifluoroacetic acid. The protecting groups may beremoved at any convenient stage in the synthesis using conventionaltechniques well known in the chemical art, or they may be removed duringa later reaction step or during work-up.

Compounds of Formulae (I), (Ia), (II), (III), (IV) or (V) may formstable pharmaceutically acceptable acid or base salts, and in such casesadministration of a compound as a salt may be appropriate. Examples ofacid addition salts include acetate, adipate, ascorbate, benzoate,benzenesulfonate, bicarbonate, bisulfate, butyrate, camphorate,camphorsulfonate, choline, citrate, cyclohexyl sulfamate,diethylenediamine, ethanesulfonate, fumarate, glutamate, glycolate,hemisulfate, 2-hydroxyethylsulfonate, heptanoate, hexanoate,hydrochloride, hydrobromide, hydroiodide, hydroxymaleate, lactate,malate, maleate, methanesulfonate, meglumine, 2-naphthalenesulfonate,nitrate, oxalate, pamoate, persulfate, phenylacetate, phosphate,diphosphate, picrate, pivalate, propionate, quinate, salicylate,stearate, succinate, sulfamate, sulfanilate, sulfate, tartrate, tosylate(p-toluenesulfonate), trifluoroacetate, and undecanoate. Examples ofbase salts include ammonium salts; alkali metal salts such as sodium,lithium and potassium salts; alkaline earth metal salts such asaluminum, calcium and magnesium salts; salts with organic bases such asdicyclohexylamine salts and N-methyl-D-glucamine; and salts with aminoacids such as arginine, lysine, ornithine, and so forth. Also, basicnitrogen-containing groups may be quaternized with such agents as: loweralkyl halides, such as methyl, ethyl, propyl, and butyl halides; dialkylsulfates such as dimethyl, diethyl, dibutyl; diamyl sulfates; long chainhalides such as decyl, lauryl, myristyl and stearyl halides; arylalkylhalides such as benzyl bromide and others. Non-toxicphysiologically-acceptable salts are preferred, although other salts maybe useful, such as in isolating or purifying the product.

The salts may be formed by conventional means, such as by reacting thefree base form of the product with one or more equivalents of theappropriate acid in a solvent or medium in which the salt is insoluble,or in a solvent such as water, which is removed in vacuo or by freezedrying or by exchanging the anions of an existing salt for another anionon a suitable ion-exchange resin.

Compounds of Formulae (I), (Ia), (II), (III), (IV) or (V) have one ormore chiral centers, and it is to be understood that the inventionencompasses all such stereoisomers, including enantiomers anddiastereoisomers. Thus, it is to be understood that, insofar as certainof the compounds of Formulae (I), (Ia), (II), (III), (IV) or (V) mayexist in optically active or racemic forms by virtue of one or moreasymmetric carbon atoms, the invention includes in its definition anysuch optically active or racemic form which possesses theabove-mentioned activity. The present invention encompasses all suchstereoisomers having activity as herein defined.

The synthesis of optically active forms may be carried out by standardtechniques of organic chemistry well known in the art, for example bysynthesis from optically active starting materials or by resolution of aracemic form. Racemates may be separated into individual enantiomersusing known procedures (see, for example, Advanced Organic Chemistry:3rd Edition: author J March, p 104-107). A suitable procedure involvesformation of diastereomeric derivatives by reaction of the racemicmaterial with a chiral auxiliary, followed by separation, for example bychromatography, of the diastereomers and then cleavage of the auxiliaryspecies. Similarly, the above-mentioned activity may be evaluated usingthe standard laboratory techniques referred to hereinafter.

Thus, throughout the specification, where reference is made to thecompound of Formulae (I), (Ia), (II), (III), (IV) or (V), it is to beunderstood that the term compound includes isomers, mixtures of isomers,and stereoisomers that are β-lactamase inhibitors.

Stereoisomers may be separated using conventional techniques, e.g.chromatography or fractional crystallisation. The enantiomers may beisolated by separation of a racemate for example by fractionalcrystallisation, resolution or HPLC. The diastereoisomers may beisolated by separation by virtue of the different physical properties ofthe diastereoisomers, for example, by fractional crystallisation, HPLCor flash chromatography. Alternatively particular stereoisomers may bemade by chiral synthesis from chiral starting materials under conditionswhich will not cause racemisation or epimerisation, or byderivatisation, with a chiral reagent.

When a specific stereoisomer is provided (whether provided byseparation, by chiral synthesis, or by other methods) it is favorablyprovided substantially isolated from other stereoisomers of the samecompound. In one aspect, a mixture containing a particular stereoisomerof a compound of Formulae (I), (Ia), (II), (III), (IV) or (V) maycontain less than 30%, particularly less than 20%, and more particularlyless than 10% by weight of other stereoisomers of the same compound. Inanother aspect, a mixture containing a particular stereoisomer of acompound of Formulae (I), (Ia), (II), (III), (IV) or (V) may containless than 6%, particularly less than 3%, and more particularly less than2% by weight of other stereoisomers of the compound. In another aspect,a mixture containing a particular stereoisomer of a compound of Formulae(I), (Ia), (II), (III), (IV) or (V) may contain less than 1%,particularly less than 0.5%, and more particularly less than 0.3%, andstill more particularly less 0.1% by weight of other stereoisomers ofthe compound.

It is to be understood that, insofar as certain of the compounds ofFormulae (I), (Ia), (II), (III), (IV) or (V) defined above may exist intautomeric forms, the invention includes in its definition any suchtautomeric form which possesses the above-mentioned activity. Thus, theinvention relates to all tautomeric forms of the compounds of Formulae(I), (Ia), (II), (III), (IV) or (V) whether explicitly detailed in thespecification or not.

It is also to be understood that certain compounds of Formulae (I),(Ia), (II), (III), (IV) or (V) and pharmaceutically salts thereof, canexist in solvated as well as unsolvated forms such as, for example,hydrated forms. It is to be understood that the invention encompassesall such solvated forms. For the sake of clarity, this includes bothsolvated (e.g., hydrated) forms of the free form of the compound, aswell as solvated (e.g., hydrated) forms of the salt of the compound.

For the sake of clarity, it should be understood that the atoms of thecompounds of Formulae (I), (Ia), (II), (III), (IV) or (V) and of any ofthe examples or embodiments disclosed herein, are intended to encompassall isotopes of the atoms. For example, H (or hydrogen) includes anyisotopic form of hydrogen including ¹H, ²H (D), and ³H (T); C includesany isotopic form of carbon including ¹²C, ¹³C, and ¹⁴C; O includes anyisotopic form of oxygen including ¹⁶O, ¹⁷O and ¹⁸O; N includes anyisotopic form of nitrogen including ¹³N, ¹⁴N and ¹⁵N; P includes anyisotopic form of phosphorous including ³¹P and ³²P; S includes anyisotopic form of sulfur including ³²S and ³⁵S; F includes any isotopicform of fluorine including ¹⁹F and ¹⁸F; Cl includes any isotopic form ofchlorine including ³⁵Cl, ³⁷Cl and ³⁶Cl; and the like. In one aspect, thecompounds of Formulae (I), (Ia), (II), (III), (IV) or (V) includeisotopes of the atoms covered therein in amounts corresponding to theirnaturally occurring abundance. However, in certain instances, it may bedesirable to enrich one or more atom in a particular isotope which wouldnormally be present in a lower abundance. For example, ¹H would normallybe present in greater than 99.98% abundance; however, in one aspect, acompound of the invention may be enriched in ²H or ³H at one or morepositions where H is present. In another aspect, when a compound of theinvention is enriched in a radioactive isotope, for example ³H and ¹⁴C,the compound may be useful in drug and/or substrate tissue distributionassays. It is to be understood that the invention encompasses all suchisotopic forms which are useful for treating bacterial infections.

In one aspect, the terms “infection” and “bacterial infection” may referto a gynecological infection. In another aspect the terms “infection”and “bacterial infection” may refer to a respiratory tract infection(RTI). In still another, the terms “infection” and “bacterial infection”may refer to a sexually transmitted disease. In yet another aspect, theterms “infection” and “bacterial infection” may refer to a urinary tractinfection (UTI). In a further aspect, the terms “infection” and“bacterial infection” may refer to acute exacerbation of chronicbronchitis (ACEB). In yet a further aspect, the terms “infection” and“bacterial infection” may refer to acute otitis media. In one aspect,the terms “infection” and “bacterial infection” may refer to acutesinusitis. In another aspect, the terms “infection” and “bacterialinfection” may refer to an infection caused by drug resistant bacteria.In still another aspect, the terms “infection” and “bacterial infection”may refer to catheter-related sepsis. In yet another aspect, the terms“infection” and “bacterial infection” may refer to chancroid. In afurther aspect, the terms “infection” and “bacterial infection” mayrefer to chlamydia. In still a further aspect, the terms “infection” and“bacterial infection” may refer to community-acquired pneumonia (CAP).In yet a further aspect, the terms “infection” and “bacterial infection”may refer to complicated skin and skin structure infection. In oneaspect, the terms “infection” and “bacterial infection” may refer touncomplicated skin and skin structure infection. In another aspect, theterms “infection” and “bacterial infection” may refer to endocarditis.In still another aspect, the terms “infection” and “bacterial infection”may refer to febrile neutropenia. In yet another aspect, the terms“infection” and “bacterial infection” may refer to gonococcalcervicitis. In a further aspect, the terms “infection” and “bacterialinfection” may refer to gonococcal urethritis. In still a furtheraspect, the terms “infection” and “bacterial infection” may refer tohospital-acquired pneumonia (HAP). In yet another aspect, the terms“infection” and “bacterial infection” may refer to osteomyelitis. In afurther aspect, the terms “infection” and “bacterial infection” mayrefer to sepsis. In still a further aspect, the terms “infection” and“bacterial infection” may refer to syphilis. In a further aspect, theterms “infection” and “bacterial infection” may refer to anintra-abdominal infection (IAI).

In one embodiment of the invention, the terms “infection” and “bacterialinfection” refer to a infection caused by Gram-negative bacteria, alsoreferred to as a “Gram-negative infection”. In one aspect of thisembodiment, the Gram-negative infection is a an infection resistant toone or more antibiotics. In one aspect of this embodiment, theGram-negative infection is a multi-drug resistant infection.

All the above mentioned infections can be caused by a variety ofbacteria that potentially could be treatable with the claimed agents incombination with penicillin-binding protein inhibitors, or by itself. Inone embodiment of the invention is a method of treating one or more ofthe infections listed above comprising administering to a subjectsuffering from a bacterial infection an effective amount of a compoundof Formulae (I), (Ia), (II), (III), (IV) or (V) or a pharmaceuticallyacceptable salt thereof, in combination with an additional antibioticagent. In one aspect of this embodiment, the additional antibiotic agentis a β-lactam antibiotic. In one aspect of this embodiment, theadditional antibiotic agent is a penicillin-binding protein inhibitor.

In one aspect, there is provided the use of a compound of Formulae (I),(Ia), (II), (III), (IV) or (V), or a pharmaceutically acceptable saltthereof, in the manufacture of a medicament for the production of abacterial peptidoglycan inhibitory effect, either alone or incombination with a penicillin-binding protein inhibitor, in awarm-blooded animal such as man.

In another aspect, there is provided the use a compound of Formulae (I),(Ia), (II), (III), (IV) or (V), or a pharmaceutically acceptable saltthereof, in the manufacture of a medicament for the treatment of abacterial infection in a warm-blooded animal such as man. In one aspect,the compound of Formulae (I), (Ia), (II), (III), (IV) or (V), or apharmaceutically acceptable salt thereof, is administered in combinationwith an additional antibiotic agent, such as a β-lactam antibiotic. Inone aspect of this embodiment, the additional antibiotic agent is apenicillin-binding protein inhibitor.

In still another aspect, there is provided the use of a compound ofFormulae (I), (Ia), (II), (III), (IV) or (V), or a pharmaceuticallyacceptable salt thereof, in the manufacture of a medicament for thetreatment of urinary tract infections, pneumonia, prostatitis, skin andsoft tissue infections, sepsis, and intra-abdominal infections, in awarm-blooded animal such as man. In one aspect of this embodiment, thecompound of Formulae (I), (Ia), (II), (III), (IV) or (V) is administeredin combination with an additional antibiotic agent. In one aspect ofthis embodiment, the additional antibiotic agent is a penicillin-bindingprotein inhibitor.

In another aspect, there is provided a method for producing a bacterialpeptidoglycan inhibitory effect, either alone or in combination with apenicillin-binding protein inhibitor, in a warm-blooded animal such asman, said method comprising administering to said animal an effectiveamount of a compound of Formulae (I), (Ia), (II), (III), (IV) or (V), ora pharmaceutically acceptable salt thereof.

In a further aspect, there is provided a method for treating a bacterialinfection in a warm-blooded animal such as man, said method comprisingadministering to said animal an effective amount of a compound ofFormulae (I), (Ia), (II), (III), (IV) or (V), or a pharmaceuticallyacceptable salt thereof. In one aspect of this embodiment, the compoundof Formulae (I), (Ia), (II), (III), (IV) or (V), or a pharmaceuticallyacceptable salt thereof, is administered in combination with anadditional antibiotic agent. In one aspect of this embodiment, theadditional antibiotic agent is a penicillin-binding protein inhibitor.In one aspect, the additional antibiotic agent is a β-lactam antibiotic.

In still a further aspect, there is provided a method for treatingurinary tract infections, pneumonia, prostatitis, skin and soft tissueinfections, sepsis, and intra-abdominal infections, in a warm-bloodedanimal such as man, said method comprising administering to said animalan effective amount of a compound of Formulae (I), (Ia), (II), (III),(IV) or (V), or a pharmaceutically acceptable salt thereof. In oneaspect of this embodiment, the compound of Formulae (I), (Ia), (II),(III), (IV) or (V), or a pharmaceutically acceptable salt thereof, isadministered in combination with an additional antibiotic agent. In oneaspect of this embodiment, the additional antibiotic agent is apenicillin-binding protein inhibitor. In one aspect, the additionalantibiotic agent is a β-lactam antibiotic.

In yet a further aspect, there is provided a compound of Formulae (I),(Ia), (II), (III), (IV) or (V), or a pharmaceutically acceptable saltthereof, for use in producing a bacterial peptidoglycan inhibitoryeffect, either alone or in combination with a penicillin-binding proteininhibitor, in a warm-blooded animal such as man. In one aspect, there isprovided a compound of Formulae (I), (Ia), (II), (III), (IV) or (V), ora pharmaceutically acceptable salt thereof, for use in treatingGram-negative bacterial infections, either alone or in combination witha beta-lactam antibiotic.

In one aspect of the invention, there is provided a method of inhibitingone or more beta-lactamase enzyme comprising administering a compound ofFormulae (I), (Ia), (II), (III), (IV) or (V), or a pharmaceuticallyacceptable salt thereof, to an animal in need thereof. In a furtheraspect, the one or more beta-lactamase enzyme is a serine beta-lactamaseenzyme. In a further aspect, the one or more beta-lactamase enzyme isselected from the group consisting of Class A, Class C and Class D. In afurther aspect, the one or more beta-lactamase enzyme is a Class Aenzyme. In a further aspect, the one or more beta-lactamase enzyme is aClass C enzyme. In a further aspect, the one or more beta-lactamaseenzyme is a Class D enzyme. In a further aspect, the one or morebeta-lactamase enzyme is a Class D enzyme and one or more of Class A andC enzymes.

The beta-lactamase inhibitors of Formulae (I), (Ia), (II), (III), (IV)or (V) can be administered in combination with any β-lactam antibioticbelonging, but not limited to, the classes of clavams, carbapenems,monobactams, pencilling, and or cephalosporins, or with any othercompound susceptible to serine β-lactamases. In one aspect of theinvention, a compound of formula (I), (Ia), (II), (III), (IV) or (V) iscombined with one or more of: penicillin, methicillin, oxacillin,nafcillin, cloxacillin, dicloxacillin, flucloxacillin, temocillin,amoxicillin, ampicillin, co-amoxiclav, azlocillin, carbenicillin,ticarcillin, mezlocillin, piperacillin, cephalexin, cephalothin,CXA-101, cefazolin, cefaclor, cefuroxime, cefamandole, cefotetan,cefoxitin, ceftriaxone, cefotaxime, cefpodoxime, cefixime, ceftazidime,ceftobiprole medocaril, cefepime, cefpirome, ceftaroline, imipenem,meropenem, ertapenem, faropenem, sulopenem, doripenem, PZ-601 (ProtezPharmaceuticals), ME1036 (Forest Labs), BAL30072, MC-1, tomopenem,tebipenemn, aztreonam, tigemonam, nocardicin A, or tabtoxinine-β-lactam.In one aspect of the invention, a compound of Formulae (I), (Ia), (II),(III), (IV) or (V) is combined with meropenem, aztreonam, orceftazidime. In one aspect of the invention, a compound of Formulae (I),(Ia), (II), (III), (IV) or (V) is combined with meropenem. In one aspectof the invention, a compound of Formulae (I), (Ia), (II), (III), (IV) or(V) is combined with aztreonam. In one aspect of the invention, acompound of Formulae (I), (Ia), (II), (III), (IV) or (V) is combinedwith ceftazidime. In one aspect of the invention, a compound of Formulae(I), (Ia), (II), (III), (IV) or (V) is combined with ceftarolinefosamil.

In another aspect of the invention, the compound of Formulae (I), (Ia),(II), (III), (IV) or (V) is administered in combination with a β-lactamantibiotic and an additional antibiotic and/or an additional β-lactamaseinhibitor. In one aspect of the invention, the additional antibioticagent is selected from one of the classes of aminoglycosides,spectinomycins, macrolides, ketolides, streptogramins, oxazolidinones,tetracyclines, fluoroquinolones, coumarin antibiotics, glycopeptides,lipoglycopeptides, nitroimidazoles, ansamycins, phenicols, mupirocyn,fosfomycin, tobramycin, linezolid, daptomycin, vancomycin, and theclasess mentioned in ANTIMICROBIAL AGENTS (ASM Press, Ed: A. Bryskier(2005)).

In one aspect of the invention, the compound of Formulae (I), (Ia),(II), (III), (IV) or (V) is administered in combination with a β-lactamantibiotic and a second agent which is designed to address β-lactamresistance. In one aspect of the invention, the compound of Formulae(I), (Ia), (II), (III), (IV) or (V) is administered in combination witha β-lactam antibiotic and a second serine beta-lactamase inhibitor. Inone aspect of the invention, the second beta-lactamase inhibitor isselected from sulbactam, tazobactam, avibactam, clavulanic acid, LK-157,LK-176, SA-1-204, SA-2-13, BLI-489 (Pfizer/Wyeth), BAL0029880 andMK7655. In another aspect of the invention, the second agent designed toaddress β-lactam resistance may be a metallo-beta-lactamase (MBL)inhibitor, also known as a Class B inhibitor.

In one aspect, there is provided a compound of Formulae (I), (Ia), (II),(III), (IV) or (V), or a pharmaceutically acceptable salt thereof, foruse in treating a bacterial infection in a warm-blooded animal, such asman.

In another aspect, there is provided a compound of Formulae (I), (Ia),(II), (III), (IV) or (V), or a pharmaceutically acceptable salt thereof,for use in treating urinary tract infections, pneumonia, prostatitis,skin and soft tissue infections, sepsis and intra-abdominal infections,in a warm-blooded animal such as man.

In still another aspect, there is provided a pharmaceutical compositioncomprising a compound of Formulae (I), (Ia), (II), (III), (IV) or (V),or a pharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable carrier, diluent, or excipient.

The compositions of the invention may be in a form suitable for oral use(for example as tablets, lozenges, hard or soft capsules, aqueous oroily suspensions, emulsions, dispersible powders or granules, syrups orelixirs), for topical use (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), for administration byinhalation (for example as a finely divided powder or a liquid aerosol),for administration by insufflation (for example as a finely dividedpowder) or for parenteral administration (for example as a sterileaqueous or oily solution for intravenous, subcutaneous, intramuscular orintramuscular dosing or as a suppository for rectal dosing). In oneaspect of the invention, the compound of Formulae (I), (Ia), (II),(III), (IV) or (V), or a pharmaceutically acceptable salt thereof, isadministered intravenously. In another aspect of the invention, thecompound of Formulae (I), (Ia), (II), (III), (IV) or (V), or apharmaceutically acceptable salt thereof, is administered intravenouslyin combination with one or more other antibacterial agent. In one aspectof this embodiment, the compound of Formulae (I), (Ia), (II), (III),(IV) or (V), or a pharmaceutically acceptable salt thereof, isadministered simultaneously with one or more other antibacterial agents.

The compositions of the invention may be obtained by conventionalprocedures using conventional pharmaceutical excipients well known inthe art. Thus, compositions intended for oral use may contain, forexample, one or more coloring, sweetening, flavoring and/or preservativeagents.

Suitable pharmaceutically acceptable excipients for a tablet formulationinclude, for example, inert diluents such as lactose, sodium carbonate,calcium phosphate or calcium carbonate; granulating and disintegratingagents such as corn starch or algenic acid; binding agents such asstarch; lubricating agents such as magnesium stearate, stearic acid ortalc; preservative agents such as ethyl or propyl p-hydroxybenzoate; andanti-oxidants, such as ascorbic acid. Tablet formulations may beuncoated or coated either to modify their disintegration and thesubsequent absorption of the active ingredient within thegastrointestinal tract, or to improve their stability and/or appearance,in either case, using conventional coating agents and procedures wellknown in the art.

Compositions for oral use may be in the form of hard gelatin capsules inwhich the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules in which the active ingredient is mixed with water oran oil such as peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions generally contain the active ingredient in finelypowdered form or in the form of nano or micronized particles togetherwith one or more suspending agents, such as sodiumcarboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents such as lecithin or condensation productsof an alkylene oxide with fatty acids (for example polyoxyethylenestearate), or condensation products of ethylene oxide with long chainaliphatic alcohols, for example heptadecaethyleneoxycetanol, orcondensation products of ethylene oxide with partial esters derived fromfatty acids and a hexitol such as polyoxyethylene sorbitol monooleate,or condensation products of ethylene oxide with long chain aliphaticalcohols, for example heptadecaethyleneoxycetanol, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand a hexitol such as polyoxyethylene sorbitol monooleate, orcondensation products of ethylene oxide with partial esters derived fromfatty acids and hexitol anhydrides, for example polyethylene sorbitanmonooleate. The aqueous suspensions may also contain one or morepreservatives such as ethyl or propyl p-hydroxybenzoate; anti-oxidantssuch as ascorbic acid); coloring agents; flavoring agents; and/orsweetening agents such as sucrose, saccharine or aspartame.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil such as arachis oil, olive oil, sesame oil or coconutoil or in a mineral oil such as liquid paraffin. The oily suspensionsmay also contain a thickening agent such as beeswax, hard paraffin orcetyl alcohol. Sweetening agents such as those set out above, andflavoring agents may be added to provide a palatable oral preparation.These compositions may be preserved by the addition of an anti-oxidantsuch as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water generally contain the activeingredient together with a dispersing or wetting agent, suspending agentand one or more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients such as sweetening, flavoring and coloring agents,may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, or a mineral oil, such as for exampleliquid paraffin or a mixture of any of these. Suitable emulsifyingagents may be, for example, naturally-occurring gums such as gum acaciaor gum tragacanth, naturally-occurring phosphatides such as soya bean,lecithin, an esters or partial esters derived from fatty acids andhexitol anhydrides (for example sorbitan monooleate) and condensationproducts of the said partial esters with ethylene oxide such aspolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening, flavoring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents such asglycerol, propylene glycol, sorbitol, aspartame or sucrose, and may alsocontain a demulcent, preservative, flavoring and/or coloring agent.

The pharmaceutical compositions may also be in the form of a sterileinjectable aqueous or oily suspension, which may be formulated accordingto known procedures using one or more of the appropriate dispersing orwetting agents and suspending agents, which have been mentioned above. Asterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally-acceptable diluent or solvent,for example a solution in 1,3-butanediol.

Compositions for administration by inhalation may be in the form of aconventional pressurized aerosol arranged to dispense the activeingredient either as an aerosol containing finely divided solid orliquid droplets. Conventional aerosol propellants such as volatilefluorinated hydrocarbons or hydrocarbons may be used and the aerosoldevice is conveniently arranged to dispense a metered quantity of activeingredient.

For further information on formulation the reader is referred to Chapter25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch;Chairman of Editorial Board), Pergamon Press 1990.

The amount of active ingredient that is combined with one or moreexcipients to produce a single dosage form will necessarily varydepending upon the host treated and the particular route ofadministration. For example, a formulation intended for oraladministration to humans will generally contain, for example, from 0.5mg to 4 g of active agent compounded with an appropriate and convenientamount of excipients which may vary from about 5 to about 98 percent byweight of the total composition. Dosage unit forms will generallycontain about 1 mg to about 1000 mg of an active ingredient. For furtherinformation on Routes of Administration and Dosage Regimes the reader isreferred to Chapter 25.3 in Volume 5 of Comprehensive MedicinalChemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press1990.

In addition to the compounds of the present invention, thepharmaceutical composition of this invention may also contain or beco-administered (simultaneously, sequentially or separately) with one ormore known drugs selected from other clinically useful classes ofantibacterial agents (for example, macrolides, quinolones, β-lactams oraminoglycosides) and/or other anti-infective agents (for example, anantifungal triazole or amphotericin). These may include carbapenems, forexample meropenem or imipenem, to broaden the therapeutic effectiveness.Compounds of this invention may also contain or be co-administered withbactericidal/permeability-increasing protein (BPI) products or effluxpump inhibitors to improve activity against gram negative bacteria andbacteria resistant to antimicrobial agents.

As stated above the size of the dose required for the therapeutic orprophylactic treatment of a particular disease state will necessarily bevaried depending on the host treated, the route of administration andthe severity of the illness being treated. Preferably a daily dose inthe range of 1-50 mg/kg is employed. Accordingly, the optimum dosage maybe determined by the practitioner who is treating any particularpatient.

In addition to its use in therapeutic medicine, the compound of Formulae(I), (II), (III) or (IV) and its pharmaceutically acceptable salts arealso useful as pharmacological tools in the development andstandardization of in vitro and in vivo test systems for the evaluationof the effects of inhibitors of DNA gyrase in laboratory animals such ascats, dogs, rabbits, monkeys, rats and mice, as part of the search fornew therapeutic agents.

Compounds of Formula (I), (Ia), (II), (III), (IV) or (V) may be preparedin a variety of ways. The processes shown below illustrates a method forsynthesizing compounds of Formula (Ia) (wherein R¹, R², and R³ unlessotherwise defined, are as defined hereinabove). The reactions areperformed in solvents appropriate to the reagents and materials employedand are suitable for the transformations being effected. Also, in thedescription of the synthetic methods described below, it is to beunderstood that all proposed reaction conditions, including choice ofsolvent, reaction atmosphere, reaction temperature, duration of theexperiment and workup procedures, are chosen to be the conditionsstandard for that reaction, which should be readily recognized by oneskilled in the art. It is understood by one skilled in the art oforganic synthesis that the functionality present on various portions ofthe molecule must be compatible with the reagents and reactionsproposed. Such restrictions to the substituents, which are compatiblewith the reaction conditions, will be readily apparent to one skilled inthe art and alternate methods must then be used. The Schemes andProcesses are not intended to present an exhaustive list of methods forpreparing the compounds of Formulae (I), (Ia), (II), (III), (IV) or (V);rather, additional techniques of which the skilled chemist is aware maybe also be used for the compounds' synthesis. The claims are notintended to be limited to the structures shown in the Schemes andProcesses.

It will also be appreciated that in some of the reactions shown in theSchemes and Processes mentioned herein, it may be necessary/desirable toprotect any sensitive groups in compounds. The instances whereprotection is necessary or desirable are known to those skilled in theart, as are suitable methods for such protection. Conventionalprotecting groups may be used in accordance with standard practice (forillustration see T. W. Greene, Protective Groups in Organic Synthesis,published by John Wiley and Sons, (1991)) and as described hereinabove.

The skilled chemist will be able to use and adapt the informationcontained and referenced within the above references, and accompanyingExamples therein and also the Examples and Scheme herein, to obtainnecessary starting materials and products.

If not commercially available, the necessary starting materials for theprocedures such as those described herein may be made by procedureswhich are selected from standard organic chemical techniques, techniqueswhich are analogous to the synthesis of known, structurally similarcompounds, or techniques which are analogous to the described procedureor the procedures described in the Examples.

It is noted that many of the starting materials for synthetic methods asdescribed herein are commercially available and/or widely reported inthe scientific literature, or could be made from commercially availablecompounds using adaptations of processes reported in the scientificliterature. The reader is further referred to Advanced OrganicChemistry, 5^(th) Edition, by Jerry March and Michael Smith, publishedby John Wiley & Sons (2001), for general guidance on reaction conditionsand reagents.

In one aspect, compounds of Formulae (I), (Ia), (II), (III), (IV) or(V), or pharmaceutically acceptable salts thereof, may be prepared bythe process outlined in Scheme 1. From the Weinreb amide, compound 1,introduction of substituents at the R³ position of Formula (I), (Ia),(II), (III), (IV) or (V) may be done via a Grignard reaction, followedby the rest of the synthetic steps shown above to yield final compounds.Compounds with different substituents at the R⁴ position can besynthesized from compound 11 by N—O reduction and deallylation followedby subsequent reaction with the amine, such as alkylation or reactionwith a substituted sulphone or a substituted isocyanate. Similarly,compounds with R¹═CH₂OR, can be made from intermediate 9, using standardalkylation techniques.

An alternative means of synthesizing compounds with substitutents at R³utilizes the Baylis-Hillman product of enone followed by standardfunctional group transformations showing below, wherein the hydroxidegroup can be transformed into a leaving group, Q, which can subsequentlybe displaced by an appropriate nucleophile.

Others R³ analogs can be made through cross-coupling of correspondinghalide enone, as shown below.

The Weinreb amide, compound 1, can be made easily prepared fromcorresponding amine by through alkylation, as shown below.

Compounds with substitution at R² can be installed via Michael Addition,according to Scheme 2 below:

In any of the above-mentioned pharmaceutical compositions, processes,methods, uses, medicaments, and manufacturing features of the instantinvention, any of the alternate embodiments of the compounds of theinvention described herein also apply.

EXAMPLES

The invention will now be further described with reference to thefollowing illustrative examples in which, unless stated otherwise:

-   -   (i) temperatures are given in degrees Celsius (° C.); operations        are carried out at room temperature or ambient temperature, that        is, in a range of 18-25° C.;    -   (ii) organic solutions were dried over anhydrous magnesium        sulfate; evaporation of organic solvent was carried out using a        rotary evaporator under reduced pressure (4.5-30 mmHg) with a        bath temperature of up to 60° C.;    -   (iii) chromatography means flash chromatography on silica gel;        thin layer chromatography (TLC) was carried out on silica gel        plates;    -   (iv) in general, the course of reactions was followed by TLC or        liquid chromatography/mass spectroscopy (LC/MS) and reaction        times are given for illustration only;    -   (v) final products have satisfactory proton nuclear magnetic        resonance (NMR) spectra and/or mass spectra data;    -   (vi) yields are given for illustration only and are not        necessarily those which can be obtained by diligent process        development; preparations were repeated if more material was        required;    -   (vii) when given, NMR data is in the form of delta values for        major diagnostic protons, given in part per million (ppm)        relative to tetramethylsilane (TMS) as an internal standard,        determined at 300 MHz in DMSO-d₆ unless otherwise stated;    -   (viii) chemical symbols have their usual meanings;    -   (ix) solvent ratio was given in volume:volume (v/v) terms;    -   (x) an ISCO Combiflash refers to flash chromatography on silica        gel using Isco Combiflash® separation system: RediSep normal        phase flash column, flow rate, 30-40 ml/min;    -   (xi) the following abbreviations may have been used:        -   ACN Acetonitrile        -   BINAP 2,2′-bis(diphenylphosphino)-1,1′-binapthyl        -   Boc₂O tert-butyloxycarbonyl anhydride        -   DAST Diethylaminosulfur trifluoride        -   DCM dichloromethane        -   DIPEA/DIEA N,N-diisopropylethylamine        -   DMAc N,N-dimethylacetamide        -   DMF N,N-dimethylformamide        -   DMAP 4-dimethylaminopyridine        -   DMSO dimethylsulfoxide        -   ee enantiomeric excess        -   EtOAc/EA ethyl acetate        -   Et₂O diethyl ether        -   GC gas chromatography        -   HATU O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium            hexafluorophosphate        -   Hex hexanes        -   HPLC high-performance liquid chromatography        -   hr/h hours        -   LDA Lithium diisopropylamide        -   MeCN acetonitrile        -   MeOH methanol        -   mins/min minutes        -   o/n overnight        -   Pd₂(dba)₃ Tris(dibenzylideneacetone)dipalladium(0)        -   iPrOH i-propanol        -   rac. racemic        -   TBAF tetra-n-butylammonium fluoride        -   TEA triethylamine        -   TFA trifluoroacetic acid        -   THF tetrahydrofuran        -   TMS trimethyl silyl        -   Tosyl, Ts para-toluenesulfonyl

Example 1(2S,5R)-2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

Dowex(R) 50WX8-100, ion-exchange resin (39 g) was conditioned bystirring for 3 hours in 2N sodium hydroxide (95 mL). The resin was thenloaded into a cartridge and washed with water until pH 7. It was thenwashed with (1/1) acetone/water, followed by water again.(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 17, 0.2997 g, 0.52 mmol) was taken up in acetoneand diluted with water. The solution was loaded on the resin and elutedwith water. The fractions containing desired product were combined andlyophilized. The desired product was obtained as a light yellow solid(140 mg, 90%).

Optical rotation: (0.1 g/dL, MeOH)=−219

MS: 278 ES+ (C₈H₁₁N₃O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.78 (m, 3H); 3.20 (m, 2H); 3.96 (m, 1H);4.11 (m, 1H); 5.42 (m, 1H); 7.25 (bs, 1H); 7.51 (bs, 1H).

Route 1 Intermediate 1:(S)-2-(1-hydroxybut-3-en-2-yl)isoindoline-1,3-dione

A 2-L reaction flask containing a stir bar and sodium carbonate (1.981g, 18.69 mmol) was placed under high vacuum and dried with a heating gunfor ten minutes. Upon cooling, the flask was backfilled with nitrogen.To it was added allylpalladium chloride dimer (0.553 g, 1.53 mmol),(1R,2R)-(+)-1,2-diaminocyclohexane-N,N′-bis(2-diphenylphosphino-1-naphthoyl)(CAS 174810-09-4) (3.36 g, 4.25 mmol), and phthalimide (50 g, 339.83mmol). The flask was then purged with nitrogen for ten minutes. 1.4 Lmethylene chloride, previously degassed with a nitrogen line for tenminutes, was then added. This suspension was placed under an atmosphereof nitrogen; it was alternately stirred and sonicated over a ten-minuteperiod to facilitate solvation. At that time it was a yellow or lightorange solution containing white solid. To this mixture was added2-vinyloxirane (24.06 g, 343.23 mmol). The resulting mixture was stirredunder a nitrogen atmosphere at ambient temperature for approximately 48hours. Analysis during that time by LCMS and TLC (1:1 hexanes:ethylacetate) suggested progression of the reaction, and final analyses bythose methods suggested complete conversion of starting material to onemajor product. The reaction mixture was filtered, and the filtrate wasconcentrated under reduced pressure. The yellow, viscous fluid wasinjected onto a 330-g silica column: a minimal volume of methylenechloride was used to thin the crude material. Silica gel chromatography(15-75% ethyl acetate in hexanes, 40 minutes, 330-g column) was used toisolate the desired product as a viscous yellow fluid that became a paleyellowish white solid (69.6 g, 94%) over a period of hours under reducedpressure.

Optical Rotation: (2.02 g/100 mL, methylene chloride) literaturevalue=−72.2, obtained value=−71.

¹H NMR (300 MHz, DMSO-d₆) δ: 3.66 (ddd, J=11.00, 6.47, 5.76 Hz, 1H) 3.97(ddd, J=10.95, 9.63, 5.67 Hz, 1H) 4.69-4.79 (m, 1H) 5.01 (dd, J=6.52,5.76 Hz, 1H) 5.18 (dt, J=2.79, 1.35 Hz, 1H) 5.23 (dt, J=9.44, 1.42 Hz,1H) 6.07 (ddd, J=17.28, 10.67, 6.42 Hz, 1H) 7.86 (q, J=1.83 Hz, 4H)

Intermediate 2:(S)-2-(1-(tert-butyldimethylsilyloxy)but-3-en-2-yl)isoindoline-1,3-dione

To a stirred solution of(S)-2-(1-hydroxybut-3-en-2-yl)isoindoline-1,3-dione (Intermediate 1,69.4 g, 319.49 mmol) and imidazole (26.1 g, 383.39 mmol) in methylenechloride (160 mL), at ambient temperature under an atmosphere ofnitrogen, was added tert-butyldimethylchlorosilane (55.4 g, 367.41 mmol)as a solid. This addition was performed over approximately ten minutes.Warming of the mixture was observed during this addition. After twohours stirring, the solution was poured into a saturated solution ofaqueous sodium bicarbonate (approximately 150 mL); this biphasic mixturewas shaken, and the organic layer was separated. The aqueous layer wasback-extracted three times with 200 mL methylene chloride each time. Theorganic layers were combined, dried over magnesium sulfate, filtered,and concentrated in vacuo. The desired product was obtained as a paleyellow solid after drying overnight under high vacuum (107 g, 101%).

¹H NMR (300 MHz, DMSO-d₆) δ: −0.13 (s, 3H) −0.04 (s, 3H) 0.67 (s, 9H)3.85 (dd, J=10.20, 5.85 Hz, 1H) 4.08 (t, J=10.01 Hz, 1H) 4.76-4.86 (m,1H) 5.20-5.33 (m, 1H) 5.25 (dt, J=2.79, 1.35 Hz, 1H) 6.08 (ddd, J=17.23,10.53, 6.42 Hz, 1H) 7.86 (dq, J=4.51, 2.21 Hz, 4H).

Intermediate 3: (S)-1-(tert-butyldimethylsilyloxy)but-3-en-2-amine

To a stirred solution of(S)-2-(1-(tert-butyldimethylsilyloxy)but-3-en-2-yl)isoindoline-1,3-dione(Intermediate 2, 108.28 g, 326.65 mmol) in methanol (1000 ml), atambient temperature under a nitrogen atmosphere, was added hydrazine(35.9 ml, 1143.29 mmol). The yellow solution was heated to 65° C. Within30 minutes of reaching reaction temperature, a white precipitate wasobserved in the reaction mixture; this solid quickly became the bulk ofthe mixture, and at that time water (about 150 mL) was added to thereaction mixture. The reaction continued stirring without interruptionand within a few minutes the solid dissolved. Upon complete conversionas indicated by LCMS analysis (both starting material and product givestrong UV signals and are easily identified by LCMS), the heat wasremoved and more water was added (a total water content of 600 mL). Themixture was allowed to come to ambient temperature.

The methanol was removed in vacuo at 35° C. (moderately reducedpressure); vacuum was removed and the aqueous was warmed to about 50° C.and then extracted with 4×200-mL methylene chloride. This approach canlead to difficulty in separation of water from organic, so plenty ofbrine should be used as the last step of the workup. The organicextracts were combined, washed with saturated sodium bicarbonate (aq),washed with brine, dried over sodium sulfate, filtered, and concentratedin vacuo at not more than 30° C. The desired product was obtained as ayellow liquid (58.5 g, 94%).

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H) 0.86 (s, 9H) 1.51 (br. s., 2H)3.22-3.30 (m, 1H) 3.33-3.48 (m, 2H) 4.98-5.05 (m, 1H) 5.17 (dt, J=17.28,1.84 Hz, 1H) 5.79 (ddd, J=17.37, 10.39, 5.67 Hz, 1H).

Intermediate 4: 2-bromo-N-methoxy-N-methylacetamide

A stirred solution of potassium carbonate (343 g, 2.48 mol) in water(about 800 mL) was prepared and cooled in an ice bath for 15 minutesunder nitrogen. To it was added O,N-dimethylhydroxylamine hydrochloride(110 g, 1.13 mol) and diethyl ether (about 800 mL). To this mixture wasthen added bromoacetyl bromide (273 g, 1.35 mol) by addition funnel overtwenty minutes. The ice bath was removed and the mixture was stirredunder nitrogen for two hours. The layers were separated and the aqueouslayer was extracted with ether (about 350 mL). The organic layers werecombined, dried over magnesium sulfate, filtered, and concentrated invacuo. The desired product was obtained as a yellow liquid (143 g, 70%).

¹H NMR (300 MHz, CDCl₃): δ 3.24 (s, 3H); 3.80 (s, 3H); 4.01 (s, 2H).

Intermediate 5: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate

A suspension of (S)-1-(tert-butyldimethylsilyloxy)but-3-en-2-amine(Intermediate 3, 60.4 g, 300 mmol) and cesium carbonate (103 g, 315mmol) in acetonitrile (about 700 mL) and water (about 120 mL) wasprepared and stirred in an ice bath under nitrogen for 5 minutes. Themixture was biphasic and remained so for the duration of the reaction.To this mixture was then added 2-bromo-N-methoxy-N-methylacetamide(Intermediate 4, 57.0 g, 285 mmol) by addition funnel over 10 minutes.The mixture was stirred for two days, with the temperature maintainednear 0° C. The mixture was kept in the freezer overnight. Analysis byTLC (ethyl acetate, potassium permanganate stain, starting amineR_(f)˜0.25) indicated high but incomplete conversion of starting amine.Another 0.05 eq of the electrophile was added. The starting amine neverdisappeared completely by TLC.

To the mixture was added di-tert-butyl dicarbonate (165 mL, 2M solutionin THF); the mixture was stirred until analysis by TLC (ethyl acetate,potassium permanganate stain) indicated consumption of intermediate. Theorganic layer was separated from the aqueous (TLC indicated that noproduct remained in the aqueous), and the organic layer was concentratedin vacuo. Silica gel chromatography (5-55% ethyl acetate in hexanes),split into 3 batches, afforded the desired product as a pale yellow oil(80 g, 66%).

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (d, J=5.10 Hz, 6H) 0.84 (s, 9H) 1.33(s, 6H) 1.38 (s, 3H) 3.02-3.15 (m, 3H) 3.61-3.68 (m, 3H) 3.70-3.86 (m,2H) 3.95-4.12 (m, 2H) 4.23-4.68 (m, 1H) 5.08-5.31 (m, 2H) 5.75-5.96 (m,1H).

Intermediate 6: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(3-methyl-2-oxobut-3-enyl)carbamate

To a solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 5, 30.79 g, 76.48 mmol) in THF (200 mL) at 0° C. was addedprop-1-en-2-ylmagnesium bromide (0.5M in THF) (300 mL, 149.90 mmol). Thereaction mixture was stirred at 0° C. for 1 hour. The reaction mixturewas quenched with 200 mL 10% citric acid, diluted further with 100 mLwater and extracted with ether. The organics were concentrated and theresulting oil was dissolved in ether and washed with water and brine.The organics were dried over magnesium sulfate, filtered andconcentrated. Silica gel chromatography (0%-20% ethyl acetate/hexanes)afforded the desired product (26.2 g, 89%) as a colorless oil.

MS: 384 ES+ (C₂₀H₃₇NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (d, 6H); 0.83 (s, 9H); 1.27-1.38 (m,9H); 1.80 (m, 3H); 3.71 (m, 2H); 4.34 (m, 2H); 4.61 (m, 1H); 5.17 (m,2H); 5.77 (m, 1H); 5.85 (m, 1H); 6.03 (m, 1H).

Intermediate 7: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

A solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(3-methyl-2-oxobut-3-enyl)carbamate(Intermediate 6, 26.18 g, 68.25 mmol) in toluene (600 mL) was purgedwith nitrogen for 15 minutes.(1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene)ruthenium(0.987 g, 1.57 mmol) was then added. The reaction mixture was heated at65° C. for 1.5 hours. The reaction mixture was concentrated onto silicagel. Silica gel chromatography (0%-15% ethyl acetate/hexanes) affordedthe desired product (21.18 g, 87%) as a colorless oil.

MS: 356 ES+ (C₁₈H₃₃NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (d, 6H); 0.81 (s, 9H); 1.42 (s, 9H);1.75 (m, 3H); 3.74-3.89 (m, 3H); 4.04-4.32 (m, 1H); 4.67 (m, 1H); 6.88(m, 1H).

Intermediate 8: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of cerium(III) chloride (14.68 g, 59.57 mmol) and(S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 7, 21.18 g, 59.57 mmol) in methanol (300 mL) at 0° C. wasadded sodium borohydride (2.254 g, 59.57 mmol) portionwise. After 15minutes, the reaction mixture was diluted with saturated ammoniumchloride (100 mL) and water (100 mL), then extracted twice with diethylether. The organic extracts were washed with brine, dried over magnesiumsulfate, filtered and concentrated. Silica gel chromatography (0%-20%ethyl acetate/hexanes) afforded the desired product (19.45 g, 91%) as acolorless oil.

MS: 358 ES+ (C₁₈H₃₅NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (s, 6H); 0.86 (s, 9H); 1.39 (s, 9H);1.69 (m, 3H); 2.63-2.72 (m, 1H); 3.59 (m, 2H); 3.82 (m, 1H); 4.03 (m,1H); 4.21 (m, 1H); 5.04 (d, 1H); 5.38 (m, 1H).

Intermediate 9: N-(allyloxy)-2-nitrobenzenesulfonamide

To a stirred solution of O-allylhydroxylamine hydrochloride (147.05 g,1341.59 mmol) in DCM (2.5 L) at 0° C., pyridine (318 mL, 3948 mmol) wasadded followed by the addition of 2-nitrobenzene-1-sulfonyl chloride(250 g, 1128.05 mmol) portionwise as a solid. The reaction mixture wasthen stirred at the same temperature for 1 h. Completion of the reactionwas monitored by TLC. The reaction mixture was quenched with 1.5 N HCl(1 L). The organic layer was separated, washed with water (250 mL),brine (250 mL), dried over anhydrous Na₂SO₄, filtered and concentratedunder vacuum to yield the residue. The crude was purified bycrystallization using EtOAc:petroleum ether (1:3) (800 mL) and afforded202 g of the title compound as a light brown solid. The mother liquorwas concentrated and purified by silica gel column chromatography (mesh60-120) using petroleum ether:EtOAc (7:3) to yield another 19.1 g of thetitle compound as a yellow solid. The total yield is 76%.

UPLC: 257 (M-1) for C₉H₁₀N₂O₅S

¹HNMR (400 MHz, DMSO-d₆): δ 4.36-4.38 (m, 2H), 5.22-5.32 (m, 2H),5.84-5.91 (m, 1H), 7.92-7.96 (m, 2H), 8.02-8.05 (m, 2H), 11.07 (s, 1H).

Intermediate 10: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 8, 19.45 g, 54.40 mmol) in toluene (300 mL) at roomtemperature was added triphenylphosphine (17.06 g, 65.28 mmol),N-(allyloxy)-2-nitrobenzenesulfonamide (Intermediate 9, 14.05 g, 54.40mmol) and diisopropyl azodicarboxylate (12.85 mL, 65.28 mmol). After 2hours the reaction mixture was concentrated onto silica gel andpurified. Silica gel chromatography (0%-50% ethyl acetate/hexanes)afforded the desired product (25.2 g, 78%) as a yellow oil.

MS: 598 ES+ (C₂₇H₄₃N₃O₈SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.00 (s, 6H); 0.83 (s, 9H); 1.31 (m, 9H);1.34 (m, 3H); 3.10-3.25 (m, 1H); 3.59 (m, 2H); 3.99-4.41 (m, 5H); 5.17(m, 2H); 5.72 (m, 2H); 7.93-8.16 (m, 4H).

Intermediate 11:N-(allyloxy)-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-4-methyl-1,2,3,6-tetrahydropyridin-3-yl)-2-nitrobenzenesulfonamide

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 10, 13.38 g, 22.38 mmol) in DCM (200 mL) at roomtemperature was added zinc bromide (3.36 mL, 67.15 mmol). The reactionmixture was stirred at room temperature overnight then diluted with DCMand washed with saturated sodium bicarbonate and brine. The organiclayer was dried over magnesium sulfate, filtered and concentrated toafford the desired product as an orange oil (11.14 g, 100%).

MS: 498 ES+ (C₂₂H₃₅N₃O₆SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (s, 6H); 0.85 (s, 9H); 1.61 (m, 3H);2.67 (m, 1H); 2.81 (m, 1H); 3.20 (m, 1H); 3.44 (m, 2H); 4.05 (m, 1H);4.30 (m, 1H); 4.40 (m, 1H); 5.22 (m, 2H); 5.82 (m, 2H); 7.90-8.15 (m,4H).

Intermediate 12:O-allyl-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-4-methyl-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine

To a solution ofN-(allyloxy)-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-4-methyl-1,2,3,6-tetrahydropyridin-3-yl)-2-nitrobenzenesulfonamide(Intermediate 11, 11.58 g, 23.27 mmol) and potassium carbonate (16.08 g,116.34 mmol) in acetonitrile (200 mL) at room temperature was addedbenzenethiol (11.95 mL, 116.34 mmol). After 3 hours, the reactionmixture was concentrated to dryness and DCM was added. The resultingsolids were removed by filtration. The filtrate was concentrated ontosilica gel. Silica gel chromatography (0%-100% ethyl acetate/hexanes)afforded the desired product (5.06 g, 69.6%) as an orange oil.

MS: 313 ES+ (C₁₆H₃₂N₂O₂Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H); 0.86 (s, 9H); 1.71 (m, 3H);2.17 (m, 1H); 2.81 (m, 2H); 3.11 (m, 2H); 3.43 (m, 2H); 4.09 (m, 2H);5.10-5.25 (m, 2H); 5.48 (m, 1H); 5.87-5.96 (m, 1H); 6.35 (d, 1H).

Intermediate 13:(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a solution ofO-allyl-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-4-methyl-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine(Intermediate 12, 2 g, 6.40 mmol) and N,N-diisopropylethylamine (4.46mL, 25.60 mmol) in acetonitrile (555 mL) at 0° C. was added triphosgene(0.760 g, 2.56 mmol) as a solution in acetonitrile (45 mL). Thetriphosgene solution was added via syringe pump at a rate of 0.1 mL/min.Once addition was complete the reaction was stirred at room temperatureovernight. The reaction mixture was concentrated to dryness, dilutedwith ethyl acetate, washed with water and brine, dried over magnesiumsulfate, filtered and concentrated. The aqueous washes were found tocontain some product and were extracted twice with ethyl acetate. Thecombined organics were dried over magnesium sulfate, filtered andcombined with previous extracts. Silica gel chromatography (0%-20% ethylacetate/hexanes) afforded the desired product (1.980 g, 91%) as a lightorange oil.

MS: 339 ES+ (C₁₇H₃₀N₂O₃Si)

¹H NMR (300 MHz, CDCl₃) δ: 0.07 (s, 6H); 0.89 (s, 9H); 1.87 (m, 3H);3.24 (m, 1H); 3.41 (m, 1H); 3.61 (m, 1H); 3.86 (m, 3H); 4.43 (m, 2H);5.33 (m, 3H); 6.01 (m, 1H).

Intermediate 14:(2S,5R)-6-(allyloxy)-2-(hydroxymethyl)-4-methyl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a solution of(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 13, 1.76 g, 5.20 mmol) in THF (20 mL) at 0° C. was addedtetrabutylammonium fluoride (1M in THF) (6.76 mL, 6.76 mmol). Stirred at0° C. for 1 hour. The reaction mixture was concentrated onto silica gel.Silica gel chromatography (50%-100% ethyl acetate/hexanes) afforded thedesired product (1.070 g, 92%) as a colorless oil.

MS: 225 ES+ (C₁₁H₁₆N₂O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.77 (m, 3H); 3.03 (m, 1H); 3.23 (m, 1H);3.49-3.60 (m, 3H); 3.78 (m, 1H); 4.36 (m, 2H); 4.84 (m, 1H); 5.24-5.39(m, 3H); 5.90-6.01 (m, 1H).

Intermediate 15:(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid

To a solution of periodic acid (2 g, 10.42 mmol) in wet acetonitrile (20mL) (0.75% water by volume) at room temperature was added chromium(VI)oxide (4 mg, 0.04 mmol). The mixture was stirred until completedissolution was achieved. This solution (5.47 mL, 3 eq) was addeddropwise at 0° C. to a solution of(2S,5R)-6-(allyloxy)-2-(hydroxymethyl)-4-methyl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 14, 0.212 g, 0.95 mmol) in wet acetonitrile (10 mL) (0.75%by volume). The reaction mixture went from clear orange to cloudybrownish color to cloudy green suspension. After 30 minutes, LC/MSshowed desired product mass and some remaining starting material.Another equivalent of the oxidizing agent solution (1.82 mL) was added.After 30 minutes, the reaction mixture was diluted with ethyl acetateand washed with 1 to 1 brine/water. The organics were dried overmagnesium sulfate, filtered and concentrated to afford a green foam(0.193 g, 86%).

MS: 239 ES+ (C₁₁H₁₄N₂O₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.80 (m, 3H); 3.19 (m, 2H); 3.85 (m, 1H);4.27 (m, 1H); 3.37 (m, 2H); 5.28-5.43 (m, 3H); 5.89-6.00 (m, 1H).

Intermediate 16:(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid (Intermediate 15, 0.193 g, 0.81 mmol) in DMF (4 mL) at roomtemperature was added ammonium chloride (0.130 g, 2.43 mmol),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (0.462 g, 1.22 mmol) and N,N-diisopropylethylamine(0.564 mL, 3.24 mmol). After 10 minutes the reaction mixture was dilutedwith ethyl acetate and washed with saturated sodium bicarbonate andbrine. The combined aqueous washes were extracted once with ethylacetate. The combined organic extracts were dried over magnesiumsulfate, filtered and concentrated. Silica gel chromatography (0%-100%ethyl acetate/hexanes) afforded a brown oil. The oil was taken up inethyl acetate and washed twice with a 1 to 1 brine/water mixture toremove DMF. The organic layer was dried over magnesium sulfate, filteredand concentrated to afford a light tan foam (0.048 g, 25%).

MS: 238 ES+ (C₁₁H₁₅N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.79 (m, 3H); 3.19 (m, 2H); 3.81 (m, 1H);4.12 (m, 1H); 4.36 (m, 2H); 5.24-5.45 (m, 3H); 5.89-6.00 (m, 1H); 7.28(bs, 1H); 7.49 (bs, 1H).

Intermediate 17: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

To a solution of(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 16, 196.9 mg, 0.83 mmol) and acetic acid (0.095 mL, 1.66mmol) (dried over sodium sulfate) in DCM (9 mL) at room temperature wasadded tetrakis(triphenylphosphine)palladium(0) (959 mg, 0.83 mmol). Thesolution was stirred at room temperature for ˜45 minutes and turned darkorange. To the reaction mixture was added pyridine (9.00 mL) and sulfurtrioxide-pyridine complex (793 mg, 4.98 mmol). The suspension wasstirred overnight at room temperature. The suspension was evaporated todryness and then resuspended in DCM. The solids were filtered offthrough a 0.45μ nalgene filter. The filtrate was concentrated to affordan orange oil. Silica gel chromatography (0%-100% acetone/DCM) affordedthe desired product (300 mg, 62.3%) as a yellow foam.

MS: 278 ES+, 303 ES+ (C₈H₁₀N₃O₆S, C₂₁H₂₀P)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.78 (m, 3H); 2.17 (m, 3H); 3.20 (m, 2H);3.96 (m, 1H); 4.11 (m, 1H); 5.42 (m, 1H); 6.57-6.74 (m, 1H); 7.22-7.30(m, 2H); 7.50 (m, 1H); 7.68-7.92 (m, 15H).

Route 2 Intermediate 18: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 10, 1 g, 1.67 mmol) in THF (11 mL) at 0° C. was addedtetrabutylammonium fluoride (1M in THF) (2.175 mL, 2.17 mmol). After 90minutes the reaction mixture was concentrated onto silica gel. Silicagel chromatography (0%-70% ethyl acetate/hexanes) afforded the desiredproduct (0.732 g, 90%) as a tan foam.

MS: 484 ES+ (C₂₁H₂₉N₃O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.31 (m, 9H); 1.35 (m, 3H); 3.20 (m, 1H);3.41 (m, 2H); 3.96-4.37 (m, 5H); 4.76 (m, 1H); 5.19 (m, 2H); 5.66-5.84(m, 2H); 7.94-8.18 (m, 4H).

Intermediate 19:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

To a solution of periodic acid (6 g, 31.26 mmol) in wet acetonitrile (60mL) (0.75% water by volume) at room temperature was added chromium(VI)oxide (10 mg, 0.10 mmol). The mixture was stirred until completedissolution was achieved.

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 18, 5 g, 10.34 mmol) in wet acetonitrile (60 mL) (0.75% byvolume) at 0° C. was added dropwise the previously formed periodicacid/chromium oxide solution (60 mL, 3 eq). After 30 minutes thereaction was complete by LC/MS. The reaction mixture was diluted withether and washed with 10% citric acid, saturated sodium bicarbonate andbrine. The organics were dried over magnesium sulfate, filtered andconcentrated to afford an orange foam (4.16 g, 81%).

MS: 498 ES+ (C₂₁H₂₇N₃O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.26 (m, 9H); 1.31 (m, 3H); 3.02-3.25 (m,1H); 3.90 (m, 1H); 4.17 (m, 3H); 4.65-4.77 (m, 1H); 5.12-5.21 (m, 2H);5.68 (m, 1H); 5.88 (m, 1H); 7.92-8.17 (m, 4H).

Intermediate 20: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 19, 4.16 g, 8.36 mmol) in DMF (35 mL) at roomtemperature was added ammonium chloride (0.895 g, 16.72 mmol), HATU(4.77 g, 12.54 mmol) and DIEA (5.84 mL, 33.45 mmol). After 15 minutesthe reaction mixture was diluted with ethyl acetate, washed withsaturated sodium bicarbonate and twice with 1:1 brine:water. Theorganics were dried over magnesium sulfate, filtered and concentrated.Silica gel chromatography (0%-80% ethyl acetate/hexanes) was run twiceto afforded the desired product (2.16 g, 52%) as a yellow foam.

MS: 497 ES+ (C₂₁H₂₈N₄O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.26 (m, 9H); 1.37 (m, 3H); 3.12-3.35 (m,1H); 3.80 (m, 1H); 4.18 (m, 3H); 4.64-4.79 (m, 1H); 5.13-5.22 (m, 2H);5.68 (m, 1H); 5.88 (m, 1H); 7.04 (m, 1H); 7.45 (bs, 1H); 7.90-8.18 (m,4H).

Intermediate 21:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 20, 2.16 g, 4.35 mmol) in DCM (20 mL) at room temperaturewas added zinc bromide (0.700 mL, 13.05 mmol). After stirring overnightat room temperature, the reaction mixture was diluted withdichloromethane and washed with saturated sodium bicarbonate and brine.The organics were dried over magnesium sulfate, filtered andconcentrated to afford the desired product (1.450 g, 84%) as a yellowfoam.

MS: 397 ES+ (C₁₆H₂₀N₄O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.65 (m, 3H); 2.71 (m, 3H); 3.76 (m, 1H);3.95 (m, 1H); 4.18-4.42 (m, 2H); 5.23 (m, 2H); 5.82 (m, 1H); 6.02 (m,1H); 7.05 (bs, 1H); 7.30 (bs, 1H); 7.93-8.18 (m, 4H).

Intermediate 22:(2S,5R)-5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamideand(2R,5R)-5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide

To a solution of(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 21, 1.4 g, 3.53 mmol) and cesium carbonate (9.21 g, 28.25mmol) in THF (100 mL) at room temperature was added PS-thiophenol(3-(3-mercaptophenyl)propanamidomethylpolystyrene) (1.55 mmol/g) (9.12g, 14.13 mmol). After stirring overnight at room temperature, thereaction mixture was filtered through a fritted funnel and the resin waswashed twice with DCM. The filtrate was concentrated to afford a yellowoil. Silica gel chromatography (0%-5% methanol/dichloromethane) affordeda 3 to 1 mixture of trans and cis isomers (0.473 g, 63.4%) as a lightyellow oil. The mixture was taken forward without separation.

MS: 212 ES+ (C₁₀H₁₇N₃O₂)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.73 (m, 3H); 2.63 (m, 1H); 2.97 (m, 1H);3.01 (m, 1H); 3.60 (m, 1H); 4.12 (m, 2H); 5.11-5.26 (m, 2H); 5.92 (m,1H); 6.45 (m, 1H); 7.00 (m, 1H); 7.33 (bs, 1H).

Intermediate 16:(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of(2S,5R)-5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamideand(2R,5R)-5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 22, 0.429 g, 2.03 mmol) and N,N-diisopropylethylamine(1.415 mL, 8.12 mmol) in acetonitrile (170 mL) at 0° C. was addedtriphosgene (0.241 g, 0.81 mmol) as a solution in acetonitrile (1.5 mL).The triphosgene solution was added at a rate of 0.1 mL/min. Onceaddition was complete the reaction was warmed to room temperature andstirred over weekend. The reaction mixture was diluted with ethylacetate, washed with saturated sodium bicarbonate and brine, dried overmagnesium sulfate, filtered and concentrated. Silica gel chromatography(0%-20% ethyl acetate/hexanes) afforded(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(0.312 g, 64.8%) as a light yellow oil.

MS: 238 ES+ (C₁₁H₁₅N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.79 (m, 3H); 3.19 (m, 2H); 3.81 (m, 1H);4.12 (m, 1H); 4.36 (m, 2H); 5.24-5.45 (m, 3H); 5.89-6.00 (m, 1H); 7.28(bs, 1H); 7.49 (bs, 1H).

The final two steps of Route 2 to yield Example 1 are equivalent forthose shown for above for Route 1.

Example 2(2S,5R)-2-cyano-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-cyano-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 24, 0.1167 g, 0.21 mmol) following the proceduredescribed for Example 1 above. The desired product was obtained as awhite solid (53.6 mg, 91%).

Optical rotation: (0.1 g/dL, DMSO)=−262

MS: 258 ES− (C₈H₉N₃O₅S)

¹H NMR (600 MHz, DMSO-d₆) δ: 1.82 (s, 3H); 3.26 (m, 1H); 3.44 (m, 1H);4.08 (m, 1H); 4.95 (m, 1H); 5.33 (m, 1H).

Intermediate 23:(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carbonitrile

To a solution of(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 16, 146 mg, 0.62 mmol) in DCM (6 mL), under nitrogen, atroom temperature was added methoxycarbonylsulfamoyl)triethyl-ammoniumhydroxide inner salt (Burgess Reagent) (660 mg, 2.77 mmol) portionwiseover 2 hours. The reaction was stirred at room temperature for anadditional 30 minutes. The reaction mixture was washed with 1:1brine:water. The organic layer was dried over magnesium sulfate,filtered and concentrated. Silica gel chromatography (0%-25% ethylacetate/hexanes) afforded(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carbonitrile(112 mg, 83%) as a colorless oil.

MS: 220 ES+ (C₁₁H₁₃N₃O₂)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.83 (m, 3H); 3.27 (m, 1H); 3.40 (m, 1H);3.97 (m, 1H); 4.38 (m, 2H); 4.97 (m, 1H); 5.26-5.39 (m, 3H); 5.88-5.99(m, 1H).

Intermediate 24:(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carbonitrile

The title compound was prepared from(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carbonitrile(Intermediate 23, 111.8 mg, 0.51 mmol) following the procedure describedfor Intermediate 17. The desired product was obtained as an off-whitefoam (117 mg, 40.8%).

MS: 258 ES−, 303 ES+ (C₈H₈N₃O₅S, C₂₁H₂₀P)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.83 (m, 3H); 2.17 (m, 3H); 3.26 (m, 1H);3.45 (m, 1H); 4.08 (m, 1H); 4.94 (m, 1H); 5.33 (m, 1H); 6.59-6.72 (m,1H); 7.22-7.39 (m, 1H); 7.68-7.92 (m, 15H).

Example 3(2S,5R)-4-methyl-7-oxo-2-(piperidinium-4-ylcarbamoyl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

To a solution of (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(1-(tert-butoxycarbonyl)piperidin-4-ylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 26, 0.1521 g, 0.20 mmol) in DCM (3 mL) at 0° C.was added trifluoroacetic acid (0.031 mL, 0.40 mmol). After 30 minutesmore trifluoroacetic acid (0.031 mL, 0.40 mmol) was added. After another30 minutes, another 6 equivalents of TFA was added at 0° C. Reactionmixture was allowed to warm to room temperature. After 1 hour, thereaction was not complete. The reaction mixture was stored in thefreezer overnight. In the morning, another 5 eq TFA were added at 0° C.Reaction allowed to warm to room temperature. After 4 hours, thereaction mixture was concentrated and coevaporated with DCM three times.The sticky oil was then triturated with ether and concentrated to afforda yellow solid. The solid was dissolved in water and washed twice withDCM. The aqueous phase was lyophilized. Purification was done on reversephase HPLC (0-10% methanol in water, YMC Carotenoid C30, 19 mm×150 mm, 5μm) to afford the title compound as a white solid (3 mg, 4.3%).

MS: 361 ES+ (C₁₃H₂₀N₄O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.53 (m, 3H); 1.73 (m, 3H); 1.78 (m, 3H);2.82 (m, 3H); 3.16 (m, 2H); 3.77 (m, 1H); 3.97 (m, 1H); 4.14 (m, 1H);5.37 (m, 1H); 6.49 (m, 1H); 8.13 (m, 1H).

Route 1 Intermediate 25: tert-butyl4-((2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-enecarboxamido)piperidine-1-carboxylate

To a solution of(2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid (Intermediately, 0.407 g, 1.71 mmol) in DMF (7 mL) at roomtemperature was added 1-Boc-4-amino-piperidine hydrochloride (0.809 g,3.42 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (1.299 g, 3.42 mmol) and N,N-diisopropylethylamine(1.190 mL, 6.83 mmol). After 30 minutes, the reaction mixture wasdiluted with ethyl acetate and washed with saturated sodium bicarbonate,brine and 1/1 brine/water. The organics were dried over magnesiumsulfate, filtered and concentrated. Silica gel chromatography (0%-60%ethyl acetate/hexanes) afforded the desired product (0.502 g, 69.9%) asan off-white foam.

MS: 421 ES+ (C₂₁H₃₂N₄O₅)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.32 (m, 2H); 1.39 (s, 9H); 1.66 (m, 2H);1.79 (m, 3H); 2.81 (m, 2H); 3.14 (m, 1H); 3.32 (m, 1H); 3.73 (m, 1H);3.84 (m, 3H); 4.14 (m, 1H); 4.36 (m, 2H); 5.24-5.42 (m, 3H); 5.88-6.02(m, 1H); 8.00 (m, 1H).

Intermediate 26: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(1-(tert-butoxycarbonyl)piperidin-4-ylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

To a solution of tert-butyl4-((2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-enecarboxamido)piperidine-1-carboxylate(Intermediate 25, 0.502 g, 1.19 mmol) and acetic acid (0.137 mL, 2.39mmol) (dried over sodium sulfate) in DCM (15 mL) at room temperature wasadded tetrakis(triphenylphosphine)palladium(0) (0.690 g, 0.60 mmol). Thesolution was stirred at room temperature for 30 minutes and turned fromyellow to orange. To the reaction mixture was added pyridine (15 mL) andsulfur trioxide-pyridine complex (1.520 g, 9.55 mmol). The suspensionwas stirred overnight at room temperature. The suspension was evaporatedto dryness and then resuspended in DCM. The solids were filtered offthrough a 0.45μ nalgene filter. The filtrate was concentrated to affordan orange oil. Silica gel chromatography (0%-50% acetone/DCM) affordedthe desired product (0.152 g, 16.7%) as a white foam.

MS: 459 ES−, 303 ES+ (C₁₈H₂₇N₄O₈S, C₂₁H₂₀P)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.36 (m, 2H); 1.41 (s, 9H); 1.68 (m, 2H);1.78 (m, 3H); 2.11 (m, 2H); 2.18 (m, 2H); 2.83 (m, 2H); 3.20 (m, 1H);3.31 (m, 1H); 3.76 (m, 1H); 3.87 (m, 2H); 3.98 (m, 1H); 4.14 (m, 1H);5.40 (m, 1H); 6.59-6.74 (m, 1H); 7.24-7.33 (m, 1H); 7.69-8.01 (m, 15H).

Route 2 Intermediate 27: (2S,5R)-1-tert-butyl 2-methyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-methyl-5,6-dihydropyridine-1,2(2H)-dicarboxylate

To a solution of(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 19, 3 g, 6.03 mmol) and potassium carbonate (3.33 g,24.12 mmol) in DMF (30 mL) at room temperature was added methyl iodide(0.454 mL, 7.24 mmol). After 1 hour the reaction mixture was dilutedwith ethyl acetate and washed three times with water. The organics weredried over magnesium sulfate, filtered and concentrated. Silica gelchromatography (0%-30% ethyl acetate/hexanes) afforded the desiredproduct (2.060 g, 66.8%) as an off-white foam.

MS: 512 ES+ (C₂₂H₂₉N₃O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.27 (m, 9H); 1.71 (m, 3H); 2.98-3.19 (m,1H); 3.65 (m, 3H); 3.91 (m, 1H); 4.17 (m, 3H); 4.81-4.96 (m, 1H);5.12-5.21 (m, 2H); 5.67 (m, 1H); 5.86 (m, 1H); 7.94-8.17 (m, 4H).

Intermediate 28: (2S,5R)-methyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate

To a solution of (2S,5R)-1-tert-butyl 2-methyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-methyl-5,6-dihydropyridine-1,2(2H)-dicarboxylate(Intermediate 27, 2.06 g, 4.03 mmol) in DCM (20 mL) at room temperaturewas added zinc bromide (0.648 mL, 12.08 mmol). After stirring overnight,the reaction mixture was diluted with dichloromethane and washed withsaturated sodium bicarbonate and brine. The organics were dried overmagnesium sulfate, filtered and concentrated to afford the desiredproduct (1.650 g, 100%) as a light yellow foam.

MS: 412 ES+ (C₁₇H₂₁N₃O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.56 (m, 3H); 2.68 (m, 1H); 2.91 (m, 1H);3.61 (s, 3H); 4.01 (m, 2H); 4.28 (m, 1H); 4.48 (m, 1H); 5.18-5.28 (m,2H); 5.85 (m, 2H); 7.91-8.17 (m, 4H).

Intermediate 29: (2S,5R)-methyl5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate and(2R,5R)-methyl5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate

To a solution of (2S,5R)-methyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate(Intermediate 28, 1.65 g, 4.01 mmol) and cesium carbonate (7.84 g, 24.06mmol) in THF (100 mL) at room temperature was added PS-thiophenol(3-(3-mercaptophenyl)propanamidomethylpolystyrene) (1.55 mmol/g) (7.76g, 12.03 mmol). After 3 hours the reaction mixture was filtered and theresin was washed with DCM. The filtrate was concentrated to afford ayellow oil. Silica gel chromatography (0%-50% methanol/DCM) afforded a 1to 1 mixture of (2S,5R)-methyl5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate and(2R,5R)-methyl5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate(0.680 g, 74.9%) as an orange oil. The mixture was taken forward withoutseparation.

MS: 227 ES+ (C₁₁H₁₈N₂O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.74 (m, 6H); 2.70 (m, 3H); 3.03 (m, 4H);3.61 (s, 3H); 3.64 (s, 3H); 3.90 (m, 2H); 4.10 (m, 4H); 5.11-5.25 (m,5H); 5.60 (m, 2H); 5.91 (m, 2H); 6.25 (m, 1H); 6.41 (m, 1H).

Intermediate 30 and Intermediate 31: (2S,5R)-methyl6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylateand (2R,5R)-methyl6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylate

To a solution of (2S,5R)-methyl5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate and(2R,5R)-methyl5-(allyloxyamino)-4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate (1to 1 mixture) (Intermediate 29, 0.68 g, 3.01 mmol) andN,N-diisopropylethylamine (2.094 mL, 12.02 mmol) in acetonitrile (250mL) at 0° C. was added triphosgene (0.357 g, 1.20 mmol) as a solution inacetonitrile (3 mL). The triphosgene solution was added via syringe pumpat a rate of 1 mL/hr. Once addition was complete, the reaction waswarmed to room temperature and stirred overnight. The reaction mixturewas diluted with ethyl acetate, washed with saturated sodium bicarbonateand brine, dried over magnesium sulfate, filtered and concentrated.Silica gel chromatography (0%-60% ethyl acetate/hexanes) afforded thedesired trans product (Intermediate 30, 0.292 g, 38%) and the undesiredcis product (Intermediate 31, 0.191 g, 25%). The cis isomer can beconverted to the trans isomer by stirring in acetonitrile with 3equivalents of triethylamine for 1 hour, followed by similar work as forthe reaction mixture.

MS: 253 ES+ (C₁₂H₁₆N₂O₄) for both cis and trans

¹H NMR (300 MHz, DMSO-d₆) trans Intermediate 30 δ: 1.80 (m, 3H); 3.12(m, 1H); 3.22 (m, 1H); 3.69 (s, 3H); 3.87 (m, 1H); 4.38 (m, 3H);5.24-5.42 (m, 3H); 5.94 (m, 1H).

¹H NMR (300 MHz, DMSO-d₆) cis Intermediate 31 δ: 1.82 (m, 3H); 3.19 (m,1H); 3.34 (m, 1H); 3.64 (s, 3H); 3.89 (m, 1H); 4.34 (m, 2H); 4.61 (m,1H), 5.23-5.37 (m, 2H); 5.50 (m, 1H); 5.92 (m, 1H).

Intermediate 32:(2R,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid

To a solution of (2S,5R)-methyl6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylate(Intermediate 30, 0.479 g, 1.90 mmol) in THF (10 mL) and water (5 mL) at0° C. was added lithium hydroxide (0.045 g, 1.90 mmol). The reaction wasstirred at 0° C. for 2 hours. Another 0.5 eq lithium hydroxide wasadded. After 2 hours the reaction mixture was neutralized carefully with1N HCl at 0° C. and the THF was evaporated. The aqueous was frozen andlyophilized to afford a light orange solid (0.464 g, 103% crude).

MS: 239 ES+ (C₁₁H₁₄N₂O₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.72 (m, 3H); 2.38 (m, 1H); 3.00 (m, 1H);3.47 (m, 1H); 3.68 (m, 1H); 3.83 (m, 1H); 4.33 (m, 1H); 5.15-55 (m, 4H);5.94 (m, 1H).

Intermediate 25: tert-butyl4-((2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-enecarboxamido)piperidine-1-carboxylate

The title compound was prepared from(2R,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid (Intermediate 32) following the procedure described in Route 1 forIntermediate 25. See Route 1 for final 2 steps.

Example 4(2S,5R)-2-carbamoyl-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 44, 71.1 mg, 0.12 mmol) following the proceduredescribed for Example 1. The desired product was obtained as a lightyellow solid (31.9 mg, 83%).

Optical rotation: (0.1 g/dL, MeOH)=−212

MS: 306 ES+ (C₁₀H₁₅N₃O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.99 (m, 6H); 2.27 (m, 1H); 3.15 (m, 1H);3.26 (m, 1H); 4.12 (m, 2H); 5.38 (m, 1H); 7.26 (bs, 1H); 7.52 (bs, 1H).

Intermediate 33:(E)-2,4,6-triisopropyl-N′-(3-methylbutan-2-ylidene)benzenesulfonohydrazide

To a suspension of 2,4,6-triisopropylbenzenesulfonyl hydrazide (5.06 g,16.95 mmol) and 3-methylbutan-2-one (1.814 mL, 16.95 mmol) in ethanol(20 mL) was added 2 drops of concentrated hydrochloric acid. Thesuspension became a solution and within a minute or two a white solidbegan to precipitate. The reaction mixture was placed in the fridge for2 hours. The white precipitate was collected by filtration to afford thedesired product (4.44 g, 71%).

MS: 367 ES+ (C₂₀H₃₄N₂O₂S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.87 (d, 6H); 1.17 (m, 18H); 1.75 (s, 3H);2.31 (m, 1H); 2.90 (m, 1H); 4.24 (m, 2H); 7.19 (s, 2H); 9.96 (s, 1H).

Intermediate 34: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(4-methyl-3-methylene-2-oxopentyl)carbamate

To a suspension of(E)-2,4,6-triisopropyl-N′-(3-methylbutan-2-ylidene)benzenesulfonohydrazide(Intermediate 33, 8 g, 21.82 mmol) in hexane (65 mL) and TMEDA (6.50 mL)at −78° C. was added dropwise n-butyllithium (1.6M in hexanes) (34.1 mL,54.56 mmol). The reaction mixture turned orange and was stirred for 30minutes at −78° C., then was warmed to 0° C. and bubbling startedimmediately. The suspension became a yellow solution. After ˜15 minutesthe bubbling stopped and (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-(methoxy(methyl)-amino)-2-oxoethyl)carbamate(Intermediate 5, 4.39 g, 10.91 mmol) was added as a solution in hexane(2 mL). After ˜15 minutes LC/MS shows desired product and no remainingWeinreb amide starting material. The reaction mixture was quenched withsaturated sodium bicarbonate and extracted with ether twice. The etherextracts were dried over magnesium sulfate, filtered and concentrated toafford a yellow oil. Silica gel chromatography (0%-10% ethylacetate/hexanes) afforded the desired product (2.219 g, 49.4%) as alight yellow oil.

MS: 412 ES+ (C₂₂H₄₁NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (m, 6H); 0.82 (m, 9H); 0.98 (m, 6H);1.28-1.38 (m, 9H); 2.77 (m, 1H); 3.71 (m, 2H); 4.33 (m, 2H); 4.62 (m,1H); 5.16 (m, 2H); 5.77 (m, 2H); 6.06 (m, 1H).

Intermediate 35: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-isopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(4-methyl-3-methylene-2-oxopentyl)carbamate(Intermediate 34, 0.839 g, 2.04 mmol) following the procedure describedfor Intermediate 7, using 0.32 eq of(1,3-Bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene)ruthenium.The desired product was obtained as a colorless oil (0.667 g, 85%).

MS: 384 ES+ (C₂₀H₃₇NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.00 (m, 6H); 0.80 (s, 9H); 1.00 (m, 6H);1.42 (s, 9H); 2.77 (m, 1H); 3.85 (m, 3H); 4.26 (m, 1H); 4.69 (m, 1H);6.80 (m, 1H).

Intermediate 36: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-isopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 35, 0.667 g, 1.74 mmol) following the procedure describedfor Intermediate 8. The desired product was obtained as a colorless oil(0.464 g, 69%).

MS: 386 ES+ (C₂₀H₃₉NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (s, 6H); 0.86 (s, 9H); 0.98 (s, 6H);1.39 (s, 9H); 2.64 (m, 2H); 3.59 (m, 2H); 3.99 (m, 2H); 4.21 (m, 1H);5.04 (d, 1H); 5.36 (m, 1H).

Intermediate 37: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 36, 1.2 g, 3.11 mmol) following the procedure describedfor Intermediate 10, using 2.4 equivalents each of triphenylphosphineand diisopropylazodicarboxylate. The desired product was obtained asyellow foam (1.22 g, 62%).

MS: 626 ES+ (C₂₉H₄₇N₃O₈SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.00 (s, 6H); 0.83 (s, 9H); 1.00 (m, 6H);1.34 (m, 9H); 3.18 (m, 1H); 3.59 (m, 2H); 4.22 (m, 4H); 4.46 (m, 1H);5.18 (m, 2H); 5.73 (m, 2H); 7.91-8.18 (m, 4H).

Intermediate 38: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 37, 0.361 g, 0.58 mmol) following the procedure describedfor Intermediate 18. The desired product was obtained as a tan foam(0.257 g, 87%).

MS: 512 ES+ (C₂₃H₃₃N₃O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.93 (m, 6H); 1.35 (m, 9H); 3.13 (m, 1H);3.40 (m, 2H); 4.18 (m, 3H); 4.41 (m, 1H); 4.75 (m, 1H); 5.20 (m, 2H);5.74 (m, 2H); 7.92-8.19 (m, 4H).

Intermediate 39:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 38, 0.727 g, 1.42 mmol) following the procedure describedfor Intermediate 19. The desired product was obtained as an off-whitefoam (0.65 g, 87%).

MS: 526 ES+ (C₂₃H₃₁N₃O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.00 (m, 6H); 1.26 (m, 9H); 2.95-3.13 (m,1H); 3.95 (m, 1H); 4.10-4.34 (m, 3H); 4.77-4.91 (m, 1H); 5.19 (m, 2H);5.67-5.87 (m, 2H); 7.92-8.20 (m, 4H).

Intermediate 40: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 39, 0.65 g, 1.24 mmol) following the proceduredescribed for Intermediate 20. The desired product was obtained as anoff-white solid (0.322 g, 51%).

MS: 525 ES+ (C₂₃H₃₂N₄O₈S)

¹H NMR (300 MHz, CDCl₃) δ: 1.00 (m, 6H); 1.31 (m, 9H); 3.20 (m, 1H);4.20 (m, 3H); 4.70-4.87 (m, 1H); 5.19 (m, 2H); 5.69 (m, 1H); 5.86 (m,1H); 7.03 (m, 1H); 7.47 (m, 1H); 7.94-8.21 (m, 4H).

Intermediate 41:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 40, 0.427 g, 0.81 mmol) following the procedure describedfor Intermediate 21. The desired product was obtained as yellow foam(0.247 g, 71%).

MS: 425 ES+ (C₁₈H₂₄N₄O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.95 (m, 6H); 2.25 (m, 1H); 2.67 (m, 2H);3.81 (m, 1H); 4.07 (m, 1H); 4.30 (m, 2H); 5.24 (m, 2H); 5.83 (m, 1H);6.03 (m, 1H); 7.03 (m, 1H); 7.32 (m, 1H); 7.93-8.18 (m, 4H).

Intermediate 42:(2S,5R)-5-(allyloxyamino)-4-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 41, 0.247 g, 0.58 mmol) following the procedure describedfor Intermediate 22. The desired product was obtained as a light yellowsolid (98 mg, 71%)

MS: 240 ES+ (C₁₂H₂₁N₃O₂)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.99 (m, 6H); 2.37 (m, 1H); 3.04 (m, 1H);3.17 (m, 1H); 3.60 (m, 1H); 4.11 (m, 2H); 5.11-5.26 (m, 2H); 5.78 (m,1H); 5.86-6.00 (m, 1H); 6.31 (m, 1H); 7.01 (bs, 1H); 7.36 (bs, 1H).

Intermediate 43:(2S,5R)-6-(allyloxy)-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

The title compound was prepared from(2S,5R)-5-(allyloxyamino)-4-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(0.0981 g, 0.41 mmol) and N,N-diisopropylethylamine (Intermediate 42,0.286 mL, 1.64 mmol) following the procedure described for Intermediate16. The desired product was obtained as a light yellow oil (69 mg, 63%).

MS: 266 ES+ (C₁₃H₁₉N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.00 (m, 6H); 2.28 (m, 1H); 3.19 (m, 2H);4.02 (m, 1H); 4.14 (m, 1H); 4.37 (m, 2H); 5.28-5.42 (m, 3H); 5.90-6.01(m, 1H); 7.28 (bs, 1H); 7.51 (bs, 1H).

Intermediate 44: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from2S,5R)-6-(allyloxy)-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 43, 68.7 mg, 0.26 mmol) following the procedure describedfor Intermediate 17, using 1 equivalent oftetrakis(triphenylphosphine)-palladium. The desired product was obtainedas a yellow oil (71 mg, 45%).

MS: 306 ES+, 303 ES+ (C₁₀H₁₅N₃O₆S, C₂₁H₂₀P)

Example 5(2S,5R)-2-cyano-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-cyano-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 46, 47.2 mg, 0.08 mmol) following the proceduredescribed for Example 1. The desired product was obtained as a whitesolid (18.2 mg, 73%).

Optical rotation: (0.1 g/dL, DMSO)=−168

MS: 286 ES− (C₁₀H₁₃N₃O₅S)

¹H NMR (600 MHz, DMSO-d₆) δ: 1.00 (m, 6H); 2.30 (m, 1H); 3.20 (m, 1H);3.50 (m, 1H); 4.24 (m, 1H); 4.97 (m, 1H); 5.25 (m, 1H).

Intermediate 45:(2S,5R)-6-(allyloxy)-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carbonitrile

The title compound was prepared from2S,5R)-6-(allyloxy)-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 43, 143.9 mg, 0.54 mmol) following the procedure describedfor Intermediate 23. The desired product was obtained as a colorless oil(114 mg, 85%).

MS: 248 ES+ (C₁₃H₁₇N₃O₂)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.00 (m, 6H); 2.32 (m, 1H); 3.21 (m, 1H);3.45 (m, 1H); 4.18 (m, 1H); 4.38 (m, 2H); 4.98 (m, 1H); 5.25-5.39 (m,3H); 5.86-6.00 (m, 1H).

Intermediate 46: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-cyano-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carbonitrile(Intermediate 45, 114.1 mg, 0.46 mmol) following the procedure describedfor Intermediate 17, using 0.75 equivalents oftetrakis(triphenyl-phosphine)palladium. The desired product was obtainedas a light yellow oil (47.2 mg, 17%).

MS: 286 ES−, 303 ES+ (C₁₀H₁₂N₃O₅S, C₂₁H₂₀P)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.01 (m, 6H); 2.17 (m, 3H); 3.21 (m, 1H);3.50 (m, 1H); 4.24 (m, 1H); 4.98 (m, 1H); 5.26 (m, 1H); 6.64-6.73 (m,1H); 7.23-7.37 (m, 1H); 7.51-7.65 (m, 1H); 7.68-7.95 (m, 15H).

Example 6(2S,5R)-2-(2-aminoethylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate

To a solution of (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(2-(tert-butoxycarbonylamino)ethylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 48, 0.238 g, 0.33 mmol) in DCM (2 mL) at 0° C. wasadded TFA (2 mL). After 30 minutes the reaction mixture was concentratedto afford an orange oil. The oil was triturated with ether three timesand ethyl acetate three times to afford an orange solid. Purificationwas done on reverse phase HPLC (0-10% acetonitrile in water, YMCCarotenoid C30, 19 mm×150 mm, 5 μm) to afford the title compound as awhite solid (25.4 mg, 13%).

Optical rotation: (0.1 g/dL, DMSO)=−120.

MS: 321 ES+ (C₁₀H₁₆N₄O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.80 (m, 3H); 2.88 (m, 2H); 3.17 (m, 1H);3.24 (m, 1H); 3.39 (m, 2H); 3.99 (m, 1H); 4.21 (m, 1H); 4.48 (m, 1H);7.43 (m, 2H); 8.25 (m, 1H).

Intermediate 47: tert-butyl2-((2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-enecarboxamido)ethylcarbamate

The title compound was prepared from(2R,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid (Intermediate 15, 0.348 g, 1.46 mmol) and N-Boc-ethylenediaminehydrochloride (0.287 g, 1.46 mmol) following the procedure described forIntermediate 25. The desired product was obtained as a light pink foam(230 mg, 41%).

MS: 381 ES+ (C₁₈H₂₈N₄O₅)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.37 (s, 9H); 1.79 (m, 3H); 3.00 (m, 2H);3.16 (m, 4H); 3.82 (m, 1H); 4.14 (m, 1H); 4.37 (m, 2H); 5.24-5.47 (m,3H); 5.90-6.00 (m, 1H); 6.81 (m, 1H); 8.07 (m, 1H).

Intermediate 48: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(2-(tert-butoxy-carbonylamino)ethylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

To a solution of tert-butyl2-((2S,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-enecarboxamido)ethylcarbamate(Intermediate 47, 230 mg, 0.60 mmol) and acetic acid (0.069 mL, 1.21mmol) (dried over sodium sulfate) in DCM (9 mL) at room temperature wasadded tetrakis(triphenylphosphine)palladium(0) (349 mg, 0.30 mmol). Thesolution was stirred at room temperature for 1 hour. To the reactionmixture was added pyridine (9.00 mL) and sulfur trioxide-pyridinecomplex (577 mg, 3.63 mmol). The suspension was stirred overnight atroom temperature. The suspension was evaporated to dryness and thenresuspended in DCM. The solids were filtered off through a 0.45μ nalgenefilter. The filtrate was concentrated to afford an orange oil. This wastaken up in DCM again and filtered through a 0.45μ filter. The filtratewas concentrated to afford an orange oil. The crude material was takento the next step without purification.

MS: 419 ES−, 303 ES+ (C₁₅H₂₃N₄O₈S, C₂₁H₂₀P)

Example 7(2S,5R)-2-(methoxymethyl)-7-oxo-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(methoxymethyl)-7-oxo-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 60, 0.283 g, 0.47 mmol) following the proceduredescribed for Example 1. The desired product was obtained after reversephase HPLC purification (2-10% acetonitrile in water, Synergi Hydro RP,19 mm×150 mm, 5 μm) as a white solid (40 mg, 26%).

Optical rotation: (0.1 g/dL, MeOH)=−170.

MS: 305 ES+ (C₁₁H₁₆N₂O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.80 (s, 3H); 3.24 (m, 5H); 3.57 (m, 2H);3.82 (m, 1H); 4.48 (m, 1H); 5.02 (m, 1H); 5.43 (m, 1H); 5.57 (m, 1H).

Intermediate 49: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-oxopent-3-enyl)carbamate

To a solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 5, 32.5 g, 80.73 mmol) in THF (400 mL) under nitrogen at0° C. was added prop-1-enylmagnesium bromide (323 ml, 161.45 mmol)dropwise. The reaction mixture was stirred at 0° C. for 1 hour, thenquenched with 400 mL 10% citric acid, diluted further with 100 mL waterand extracted with ether. The organics were concentrated and theresulting oil was dissolved in ether and washed with water and brine.The organics were dried over magnesium sulfate, filtered andconcentrated. Silica gel chromatography (5%-20% ethyl acetate/hexanes)afforded the desired product as a colorless oil (27 g, 87%).

MS: 384 ES+ (C₂₀H₃₇NO₄Si)

¹H NMR (300 MHz, CDCl₃) δ: 0.05 (2, 6H); 0.88 (s, 9H); 1.39-1.47 (m,9H); 1.90 (m, 3H); 3.80 (m, 2H); 4.05-4.18 (m, 2H); 4.43-4.76 (m, 1H);5.22 (m, 2H); 5.86 (m, 1H); 6.21 (m, 1H); 6.91 (m, 1H).

Intermediate 50: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

(S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-oxopent-3-enyl)carbamate(Intermediate 49, 27.0 g, 70.39 mmol) was dissolved in toluene (650 ml).The solution was purged with nitrogen for 15 minutes before the additionof Hoveyda-Grubbs Catalyst 2nd Generation (0.885 g, 1.41 mmol). Thereaction mixture was heated under nitrogen at 65° C. After 2 hours LCMSshowed complete formation of the product. The reaction mixture wasconcentrated under reduced pressure. Silica gel chromatography (10%-35%ethyl acetate/hexanes) afforded the desired product as a solid (17.0 g,70%).

Optical Rotation: 0.1 g/dL, methylene chloride=−175

MS: 342 ES+ (C₁₇H₃₁Na₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (s, 6H); 0.82 (s, 9H); 1.43 (s, 9H);3.78-3.93 (m, 3H); 4.29 (m, 1H); 4.70 (m, 1H); 6.19 (dd, 1H); 7.15 (m,1H).

Intermediate 51: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-iodo-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 50, 10 g, 29.28 mmol) and 4-dimethylaminopyridine (0.894g, 7.32 mmol) in THF (100 mL)/water (100 mL) at room temperature wasadded potassium carbonate (3.24 g, 23.42 mmol) and iodine (8.92 g, 35.14mmol). After stirring for 15 minutes, the reaction mixture was dilutedwith ether and washed with saturated sodium thiosulfate twice, then 5%citric acid and brine. The organics were dried over magnesium sulfate,filtered and concentrated. Silica gel chromatography (0%-15% ethylacetate/hexanes) afforded the desired product as a tan oil (11.6 g,85%).

MS: 468 ES+ (C₁₇H₃₀INO₄Si)

¹H NMR (300 MHz, CDCl₃) δ: 0.04 (s, 6H); 0.86 (s, 9H); 1.49 (s, 9H);3.81 (m, 1H); 3.95 (m, 1H); 4.17 (m, 1H); 4.79 (m, 2H); 7.67 (m, 1H).

Intermediate 52: (2S,5R)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-iodo-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-iodo-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 51, 10 g, 21.39 mmol) following the procedure describedfor Intermediate 8. The desired product was obtained as a colorless oil(8.87 g, 88%).

MS: 470 ES+ (C₁₇H₃₂INO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H); 0.86 (s, 9H); 1.40 (s, 9H);2.87 (m, 1H); 3.65 (m, 2H); 3.79 (m, 1H); 4.21 (m, 2H); 5.74 (d, 1H);6.44 (m, 1H).

Intermediate 53: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(prop-1-en-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate

A solution of (2S,5R)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-iodo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 52, 8.57 g, 18.26 mmol), potassiumtrifluoro(prop-1-en-2-yl)borate (5.40 g, 36.51 mmol), potassiumcarbonate (3.11 mL, 54.77 mmol) anddichloro[1,1′-bis(di-t-butylphosphino)ferrocene]palladium(II) (1.190 g,1.83 mmol) in dioxane (200 mL) and water (66.7 mL) at room temperaturewas purged with argon for 5 minutes then heated at 70° C. The reactionmixture was concentrated onto silica gel. Silica gel chromatography(0%-20% ethyl acetate/hexanes) afforded a 2 to 1 mixture of desiredproduct and starting material as a brown oil (5.36 g, 77%).

MS: 384 ES+ (C₂₀H₃₇NO₄Si)2

Intermediate 54: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-(prop-1-en-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(prop-1-en-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 53, 3.86 g, 10.06 mmol) following the procedure describedfor Intermediate 10. The desired product was obtained as light yellowfoam (3.25 g, 52%).

MS: 624 ES+ (C₂₉H₄₅N₃O₈SSi)

Intermediate 55:N-(allyloxy)-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-4-(prop-1-en-2-yl)-1,2,3,6-tetrahydropyridin-3-yl)-2-nitrobenzenesulfonamide

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-(prop-1-en-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 54, 3.04 g, 4.87 mmol) following the procedure describedfor Intermediate 21. The desired product was obtained as an orange oil(2.53 g, 99%).

MS: 524 ES+ (C₂₄H₃₇N₃O₆SSi)

Intermediate 56:O-allyl-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-4-(prop-1-en-2-yl)-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine

The desired product was prepared fromN-(allyloxy)-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-4-(prop-1-en-2-yl)-1,2,3,6-tetrahydropyridin-3-yl)-2-nitrobenzenesulfonamide(Intermediate 55, 2.45 g, 4.68 mmol) following the procedure describedfor Intermediate 22. The desired product was obtained as yellow oil(1.19 g, 75%).

MS: 339 ES+ (C₁₈H₃₄N₂O₂Si)

Intermediate 57:(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

The title compound was prepared fromO-allyl-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)-methyl)-4-(prop-1-en-2-yl)-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine(Intermediate 56, 1.22 g, 3.60 mmol) following the procedure describedfor Intermediate 16. The desired product was obtained as a light yellowoil (1.16 g, 88%).

MS: 365 ES+ (C₁₈H₃₂N₂O₃Si)

Intermediate 58:(2S,5R)-6-(allyloxy)-2-(hydroxymethyl)-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

The title compound was obtained from(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)-methyl)-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 57, 1.16 g, 3.18 mmol) following the procedure describedfor Intermediate 14. The desired product was obtained as a colorless oil(741 mg, 93%).

MS: 251 ES+ (C₁₃H₁₈N₂O₃)

Intermediate 59:(2S,5R)-6-(allyloxy)-2-(methoxymethyl)-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a solution of(2S,5R)-6-(allyloxy)-2-(hydroxymethyl)-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 58, 0.289 g, 1.15 mmol) in DMF (10 mL) at 0° C. was addedmethyl iodide (0.435 mL, 6.93 mmol) followed by sodium hydride (60% inmineral oil) (0.051 g, 1.27 mmol). The reaction was stirred for 1.5hours at 0° C. The reaction mixture was diluted with ethyl acetate andwashed twice with water. The organics were dried over magnesium sulfate,filtered and concentrated. Silica gel chromatography (0%-50% ethylacetate/hexanes) afforded the desired product as a pale yellow oil (233mg, 76%).

MS: 265 ES+ (C₁₄H₂₀N₂O₃)

Intermediate 60: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(methoxymethyl)-7-oxo-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

The title compound was prepared from2S,5R)-6-(allyloxy)-2-(methoxymethyl)-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 59, 233 mg, 0.88 mmol) following the procedure describedfor Intermediate 17. The desired product was obtained as an off-whitefoam (283 mg, 53%).

MS: 305 ES+, 303 ES+ (C₁₁H₁₆N₂O₆S, C₂₁H₂₀P)

Example 8(2S,5R)-2-((5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

To a solution of(2S,5R)-2-((4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate (Intermediate 68, 58 mg, 0.09 mmol) in DCM (2 mL) atroom temperature was added trifluoroacetic acid (0.505 mL, 6.55 mmol).The reaction mixture was stirred for 10 minutes and then concentrated.The resulting residue was dissolved in DCM and concentrated twice more.The product was purified twice by C18 RediSepRf Gold column (15.5 g)eluting with water manually using a syringe. The desired product wasobtained after lyophilization as a white solid (3.4 mg, 9%).

MS: 401 ES+ (C₁₄H₁₆N₄O₈S)

¹H NMR (300 MHz, D₂O) δ: 1.93 (m, 3H); 3.25 (m, 1H); 3.58 (m, 1H); 4.20(m, 1H); 4.50 (m, 2H); 4.59 (m, 1H); 5.67 (m, 1H); 6.76 (s, 1H); 7.77(s, 1H).

Intermediate 61: 2-(hydroxymethyl)-5-(4-methoxybenzyloxy)-4H-pyran-4-one

To a solution of 5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one (Alfa Aesar,5.11 g, 35.96 mmol) in DMF (70 mL) at room temperature was addedpotassium carbonate (9.94 g, 71.92 mmol) and1-(chloromethyl)-4-methoxybenzene (5.86 mL, 43.15 mmol) dropwise. Thereaction mixture was heated at 80° C. for 1 hour then concentrated. Tothe resulting slurry was added ice water. The precipitate was collectedby filtration then triturated with ethyl acetate and filtered again. Thetitle compound was obtained as a tan solid (6.44 g, 68%).

MS: 263 ES+ (C₁₄H₁₄O₅)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.76 (s, 3H); 4.28 (s, 2H); 4.86 (s, 2H);5.75 (m, 1H); 6.31 (s, 1H); 6.94 (m, 2H); 7.33 (m, 2H); 8.13 (s, 1H).

Intermediate 62:2-(hydroxymethyl)-5-(4-methoxybenzyloxy)pyridin-4(1H)-one

2-(hydroxymethyl)-5-(4-methoxybenzyloxy)-4H-pyran-4-one (Intermediate63, 6.44 g, 24.56 mmol) and ammonia (7N in MeOH) (59.6 ml, 417.45 mmol)were combined in a pressure reactor vessel and heated at 90° C. for 5hours. The reaction mixture was cooled overnight then concentrated. Thesolid was suspended in water then collected by filtration. The titlecompound was obtained as a brown solid (3.48 g, 54%).

MS: 262 ES+ (C₁₄H₁₅NO₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.75 (s, 3H); 4.32 (s, 2H); 4.93 (s, 2H);5.53 (m, 1H); 6.07 (m, 1H); 6.92 (m, 2H); 7.26 (m, 1H); 7.32 (m, 2H);11.02 (m, 1H).

Intermediate 63: (4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methanol

To a solution of2-(hydroxymethyl)-5-(4-methoxybenzyloxy)pyridin-4(1H)-one (Intermediate62, 3.48 g, 13.32 mmol) in DMF (100 mL) at room temperature was added4-methoxybenzyl chloride (1.987 mL, 14.65 mmol) followed by potassiumcarbonate (2.273 mL, 39.96 mmol). The reaction mixture was stirred for 1hour at room temperature then heated at 80° C. for 1.5 hours. Thereaction mixture was cooled to room temperature, poured into water andextracted twice with ethyl acetate. The combined extracts were washedwith water and brine. The ethyl acetate layer was concentrated to affordan oil. To the oil was added 1N HCl and a light brown solid crashed out.This was collected by filtration. The solid was taken up in ethylacetate and washed with saturated sodium bicarbonate. The organics weredried over magnesium sulfate, filtered and concentrated to afford thetitle compound as a brown solid (2.62 g, 52%).

MS: 382 ES+ (C₂₂H₂₃NO₅)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.74 (s, 3H); 3.76 (s, 3H); 4.42 (m, 2H);5.05 (s, 2H); 5.12 (s, 2H); 5.27 (m, 1H); 6.93 (m, 4H); 7.17 (s, 1H);7.32 (m, 4H); 8.07 (s, 1H).

Intermediate 64:2-((4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methyl)isoindoline-1,3-dione

To a solution of (4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methanol(Intermediate 63, 1.5 g, 3.93 mmol), phthalimide (0.579 g, 3.93 mmol)and triphenylphosphine (1.028 g, 3.93 mmol) in THF (15 mL) at roomtemperature was added diisopropyl azodicarboxylate (2.091 mL, 10.62mmol). The reaction was stirred at room temperature overnight thenconcentrated onto silica gel. Silica gel chromatography (0%-70% ethylacetate) afforded the title compound as a light brown solid (1.1 g,55%).

MS: 511 ES+ (C₃₀H₂₆N₂O₆)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.73 (s, 3H); 3.74 (s, 3H); 4.78 (s, 2H);5.01 (s, 2H); 5.10 (s, 2H); 6.89 (m, 4H); 7.16 (s, 1H); 7.32 (m, 4H);7.89 (m, 4H); 8.01 (s, 1H).

Intermediate 65: (4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methanamine

To a solution of2-((4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methyl)isoindoline-1,3-dione(Intermediate 64, 1.1 g, 2.15 mmol) in chloroform (20 mL) and methanol(10 mL) at room temperature was added hydrazine hydrate (0.328 mL, 4.31mmol). The reaction was stirred overnight at room temperature. Another 1eq of hydrazine hydrate was added. After 4 hours the reaction mixturewas filtered to remove the solids. The filtrate was concentrated toafford an orange oil. The oil was dissolved in methanol and ether wasadded to crash out more solids. This was repeated until no2,3-dihydrophthalazine-1,4-dione by product remained. The title compoundwas obtained as an orange foam (0.82 g, 100%).

MS: 381 ES+ (C₂₂H₂₄N₂O₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.66 (s, 2H); 3.74 (s, 3H); 3.76 (s, 3H);5.03 (s, 2H); 5.11 (s, 2H); 6.92 (m, 4H); 7.20 (s, 1H); 7.35 (m, 4H);8.05 (s, 1H).

Intermediate 66:(2S,5R)-6-(allyloxy)-N-((4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of(R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid (Intermediate 32, 0.469 g, 1.97 mmol) in DMF (10 mL) at roomtemperature was added(4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methanamine (Intermediate 65,0.824 g, 2.17 mmol),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (1.497 g, 3.94 mmol) and N,N-diisopropylethylamine(1.372 mL, 7.87 mmol). After 30 minutes the reaction mixture was dilutedwith ethyl acetate and washed with saturated sodium bicarbonate, brine,and 1/1 brine/water twice. The organics were dried over magnesiumsulfate, filtered and concentrated. Silica gel chromatography (0%-50%ethyl acetate/hexanes) afforded the title compound as a light pink foam(0.427 g, 36%).

MS: 601 ES+ (C₃₃H₃₆N₄O₇)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.80 (m, 3H); 3.20 (m, 2H); 3.74 (s, 3H);3.76 (s, 3H); 3.80 (m, 1H); 4.28 (m, 3H); 4.38 (m, 2H); 5.05 (s, 2H);5.08 (s, 2H); 5.26 (m, 2H); 5.49 (m, 1H); 5.95 (m, 1H); 6.93 (m, 4H);7.03 (s, 1H); 7.35 (m, 4H); 8.09 (s, 1H); 8.53 (m, 1H).

Intermediate 67: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-((4,5-bis(4-methoxy-benzyloxy)pyridin-2-yl)methylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

To a solution of(2S,5R)-6-(allyloxy)-N-((4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 66, 0.427 g, 0.71 mmol) and acetic acid (0.081 mL, 1.42mmol) (dried over sodium sulfate) in DCM (10 mL) at room temperature wasadded tetrakis(triphenylphosphine)palladium(0) (0.821 g, 0.71 mmol). Thesolution was stirred at room temperature for 1 hour. To the reactionmixture was added pyridine (10.00 mL) and sulfur trioxide-pyridinecomplex (0.679 g, 4.27 mmol). The suspension was stirred overnight atroom temperature. The suspension was evaporated to dryness and thenresuspended in DCM. The solids were filtered off through a 0.45μ nalgenefilter. The filtrate was concentrated and loaded onto a 24 g RediSepsilica column through a 0.45μ nalgene filter. Silica gel chromatography(0%-100% acetone/DCM) afforded the title compound as a yellow foam(0.393 g, 59%).

MS: 639 ES−, 303 ES+ (C₃₀H₃₂N₄O₁₀S, C₂₁H₂₀P)

Intermediate 68:(2S,5R)-2-((4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The Dowex(R) 50WX8-100, ion-exchange resin (35 g) was conditioned bystirring for 3 hours in 2N sodium hydroxide (80 mL). The resin was thenloaded into a glass column (2×12 inches) and washed with water until thepH was 7. It was then washed with (1/1) acetone/water (˜500 mL),followed by water (˜500 mL). (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-((4,5-bis(4-methoxybenzyloxy)pyridin-2-yl)methylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 67, 0.393 g, 0.42 mmol) was taken up in acetone(˜2 mL) and diluted with water (˜4 mL). The yellow solution was loadedon the resin and eluted with water. The title compound was obtainedafter lyophilization as an off-white solid (124 mg, 45%).

MS: 641 ES+ (C₃₀H₃₂N₄O₁₀S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.79 (m, 3H); 3.23 (m, 2H); 3.75 (m, 6H);3.98 (m, 1H); 4.27 (m, 3H); 5.07 (m, 4H); 5.47 (m, 1H); 6.93 (m, 4H);7.03 (s, 1H); 7.34 (m, 4H); 8.10 (s, 1H).

Example 9(2S,5R)-2-carbamoyl-4-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The Dowex(R) 50WX8-100, ion-exchange resin (10 g) was conditioned bystirring for 3 hours in 2N sodium hydroxide (30 mL). The resin was thenloaded into a cartridge and washed with water until the pH was 7. It wasthen washed with (1/1) acetone/water (˜100 mL), followed by water (˜100mL). (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 76, 131 mg, mmol) was taken up in water (˜1 mL)and minimum acetonitrile. The yellow solution was loaded on the resinand washed through with water. The title compound was obtained afterlyophilization as an off-white solid (38 mg, 50%).

MS: 308 ES+ (C₉H₁₂N₃NaO₇S)

¹H NMR (300 MHz, D₂O) δ: 3.43 (d, 1H); 3.53 (s, 3H); 3.59 (m, 1H); 3.80(m, 1H); 4.24 (q, 2H); 4.45 (m, 1H); 6.11 (m, 1H).

Intermediate 69: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(hydroxymethyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(17.37 g, 46.75 mmol) in DCM (480 mL) was addedN1,N1,N8,N8-tetramethylnaphthalene-1,8-diamine (60.1 g, 280.51 mmol).Then it was cooled to 0° C. and trimethyloxonium tetrafluoroborate(20.74 g, 140.25 mmol) was added. It was then stirred at rt for 6 h.Then it was concentrated under reduced pressure. The residue was thentaken up in 100 mL Et₂O and filtered, washed with 400 mL Et₂O. Theorganic layer was then washed with 10% citric acid, aq. NaHCO₃, brine,dried over MgSO₄, filtered and concentrated to afford the desiredproduct (16.73 g, 93%) as an oil.

MS: 386 ES+ (C₁₉H₃₅NO₅Si)

¹H NMR (300 MHz, CDCl₃) δ: 0.01 (s, 6H); 0.81 (s, 9H); 1.42 (s, 9H);3.28 (s, 3H); 3.91 (m, 5H); 4.33 (d, 1H); 4.78 (m, 1H); 7.01 (s, 1H).

Intermediate 70: (2S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

(S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate)(Intermediate 69, crude, 16.73 g, 43.39 mmol) was dissolved in MeOH (100mL), cooled to 0° C. and CeCl₃ (10.69 g, 43.39 mmol) was added to give asolution. Then NaBH₄ (1.642 g, 43.39 mmol) was added slowly as solid,and the mixture was stirred from 0° C. to rt for 30 min. The volatilesolvent was removed. The white solid was redissolved in 200 mL EtOAc andwashed with sat. NaHCO₃, brine, dried over MgSO₄, filtered andconcentrated. The residue was purified by silica gel column (0-100%EA/Hex) to afford 13.78 g, 82% as an colorless oil.

MS: 410 ES+ (C₁₉H₃₇NO₅Si+Na⁺)

¹H NMR (300 MHz, CDCl₃) δ: 0.04 (s, 6H); 0.89 (s, 9H); 1.46 (s, 9H);3.10 (m, 1H); 3.36 (s, 3H); 3.68 (m, 1H); 3.72 (m, 1H); 4.11 (m, 2H);4.24 (m, 2H); 4.39 (m, 1H); 5.75 (m, 1H).

Intermediate 71: (2S,5R)-tert-butyl5-(allyloxy(tert-butoxycarbonyl)amino)-2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 70, 13.78 g, 35.55 mmol) in DCM (200 mL) at 0° C.,pyridine (14.38 mL, 177.77 mmol) and N,N-dimethylpyridin-4-amine (217mg, 1.78 mmol) was added. Then methanesulfonic anhydride (9.29 g, 53.33mmol) was added. The mixture was then stirred from 0° C. to rt for 2hrs. It was diluted with DCM (200 mL) and washed with brine, dried overMgSO₄, filtered and concentrated to give (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-5-(methylsulfonyloxy)-5,6-dihydropyridine-1(2H)-carboxylate(crude, 16.9 g) as an pale yellow oil. It was used directly for the nextstep.

To a stirred solution of tert-butyl allyloxycarbamate (7.39 g, 42.66mmol) in DMF (150 mL) at rt, potassium 2-methylpropan-2-olate (42.66 mL,42.66 mmol) was added and gave a purple solution. After 30 min at rt,the mixture was cooled to 0° C. and (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-5-(methylsulfonyloxy)-5,6-dihydropyridine-1(2H)-carboxylate(crude, 16.55 g, 35.55 mmol) in 50.0 mL DMF was added. The mixture wasthen warmed up to rt for 1 h. It was then diluted with ethyl acetate(200 mL) and washed aqueous sat. NaHCO₃ solution, brine, dried overMgSO₄, filtered and concentrated to give a residue which contains somestarting material. The crude was purified on a silica gel columnyielding a colorless oil. (19.3 g). ¹HNMR shows still a mixture (withhydroxyamine starting material). It was used directly in TBSdeprotection.

MS: 565 ES+(C₂₇H₅₀N₂O₇Si+Na⁺)

¹H NMR (300 MHz, CDCl₃) δ: 0.04 (s, 6H); 0.89 (s, 9H); 1.47 (s, 9H);1.52 (s, 9H); 3.14 (m, 1H); 3.31 (s, 3H); 3.72 (m, 3H); 4.15 (m, 3H);4.44 (m, 3H); 5.28 (m, 2H); 5.97 (m, 2H).

Intermediate 72: (2S,5R)-tert-butyl5-(allyloxy(tert-butoxycarbonyl)amino)-2-(hydroxymethyl)-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of (2S,5R)-tert-butyl5-(allyloxy(tert-butoxycarbonyl)amino)-2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 71, 35.55 mmol) in THF (100 mL) at 0° C., TBAF (39.11 mL,39.11 mmol) was added. After 1 hr at 0° C., it was then concentrated togive a residue which was purified by silica gel column (0-100% EA/Hex)to give the desired product (8.82 g, 57.9% over 3 steps) as a colorlessoil.

MS: 451 ES+ (C₂₁H₃₆N₂O₇+Na⁺)

¹H NMR (300 MHz, CDCl₃) δ: 1.47 (s, 9H); 1.52 (s, 9H); 3.14 (dd, 1H);3.34 (s, 3H); 3.71 (m, 3H); 3.98 (d, 1H), 4.18 (m, 2H); 4.40 (m, 2H);4.67 (m, 1H); 5.20 (m, 2H); 5.79 (m, 1H); 5.97 (s, 1H).

Intermediate 73: (2S,5R)-tert-butyl5-(allyloxy(tert-butoxycarbonyl)amino)-2-carbamoyl-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

Stock oxidation solution: ˜480 mg conc. HNO₃ and ˜160 mg Na₂Cr₂O₇-2H₂Owas dissolved in 32 mL H₂O at rt.

To a stirred solution of (2S,5R)-tert-butyl5-(allyloxy(tert-butoxycarbonyl)amino)-2-(hydroxymethyl)-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 72, 8.82 g, 20.58 mmol) in 100 mL MeCN at 0° C., was addedsodium periodate (19.37 g, 90.56 mmol). Then 12 mL of the stockoxidation solution was added. The suspension was then stirred at rt for2d. An additional 16 mL the above stock solution was added and stirredfor 2 more days. The mixture was diluted with 150 mL ethyl acetate, 50mL 1M pH 7 buffer, and 50 mL 2M NaHSO₃. The aqueous was extracted with20 mL ethyl acetate. The ethyl acetate layer was then washed with brine.The aqueous layer was checked by LCMS and found containing desiredcarboxylic acid. The aqueous layer was then extracted with 50 mL ethylacetate and washed with brine. The combined organic layers was driedover MgSO₄, filtered and concentrated to afford a yellow dry film,(crude 7.11 g, 78%) which was used directly without furtherpurification. To a stirred solution of(2S,5R)-5-(allyloxy(tert-butoxycarbonyl)amino)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)-1,2,5,6-tetrahydropyridine-2-carboxylicacid (crude, 7.11 g, 16.07 mmol), ammonium chloride (1.719 g, 32.14mmol), HATU (12.22 g, 32.14 mmol) in DMF (50.0 mL) at rt, was addedDIPEA (8.31 g, 64.27 mmol). After stirring at rt for 1 hr, 100 mL EtOAcwas added. The organic layer was washed with water, brine. The residuewas purified by silica gel column (0-100% Hex/EA) to afford the desiredproduct (3.07 g, 43.3%) as an off-white solid.

MS: 442 ES+ (C₂₁H₃₅N₃O₇)

¹H NMR (300 MHz, CD₃OD) δ: 1.47 (s, 9H); 1.52 (s, 9H); 3.17 (d, 1H);3.34 (s, 3H); 4.04 (m, 1H); 4.12 (m, 4H); 4.44 (m, 2H); 5.21 (m, 2H);5.77 (m, 1H); 6.22 (s, 1H).

Intermediate 74:(2S,5R)-5-(allyloxyamino)-4-(methoxymethyl)-1,2,5,6-tetrahydropyridine-2-carboxamide

(2S,5R)-tert-butyl5-(allyloxy(tert-butoxycarbonyl)amino)-2-carbamoyl-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 73, 3.07 g, 6.95 mmol) was dissolved in 20 mL DCM. Then5.36 mL TFA was added dropwise at 0° C. The mixture was stirred at rtfor 3 h. The solvent was removed in vacuo and co-evaporated twice with 5mL MeOH. The residue was dissolved in MeOH (10 mL) and ammoniumhydroxide (30% in water) was added until it was basic. The mixture wasrotovapored at rt and the residue was freeze-dried over night to give asolid. It was dissolved in DCM and purified on silica gel eluting with0-100% MeOH/DCM to give a off-white solid (1.12 g, 66.8%).

MS: 242 ES+ (C₁₁H₁₉N₃O₃)

¹H NMR (300 MHz, CD₃OD) δ: 3.31 (s, 3H); 3.55 (m, 3H); 4.05 (m, 4H);4.58 (m, 1H); 5.20 (m, 2H); 5.96 (m, 2H).

Intermediate 75:(2S,5R)-6-(allyloxy)-4-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a stirred solution of(2S,5R)-5-(allyloxyamino)-4-(methoxymethyl)-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 74, 1.12 g, 4.64 mmol) in acetonitrile (100 mL),N-ethyl-N-isopropylpropan-2-amine (4.04 mL, 3.00 g, 23.21 mmol) wasadded, then triphosgene (551 mg, 1.86 mmol) in 20 mL acetonitrile wasadded slowly via syringe pump over 4 h at 0° C. It was stirred from 0°C. to rt overnight. The solution was concentrated to give a residue,which was then taken up in 50 mL EtOAc and washed with brine, dried overMgSO₄, filtered and concentrated. The residue was purified on a silicagel column eluting with 0-100% ethyl acetate/hexanes gave a yellow oil(420 mg, 34%).

MS: 268 ES+ (C₁₂H₁₇N₃O₄)

¹H NMR (300 MHz, CD₂Cl₂) δ: 3.03 (d, 1H); 3.28 (s, 3H); 3.40 (m, 2H);3.95 (m, 3H); 4.38 (m, 2H); 5.30 (m, 2H); 5.98 (m, 2H); 6.83 (bs, 2H).

Intermediate 76: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

To a solution of(2S,5R)-6-(allyloxy)-4-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 75, 120 mg, 0.45 mmol) and acetic acid (0.051 mL, 0.90mmol) (dried over sodium sulfate) in DCM (4.0 mL) at room temperaturewas added tetrakis(triphenylphosphine)palladium(0) (519 mg, 0.45 mmol).The solution was stirred at room temperature for 1 hour. To the reactionmixture was added pyridine (2.0 mL) and sulfur trioxide-pyridine complex(429 mg, 2.69 mmol). The suspension was stirred overnight at roomtemperature. The suspension was evaporated to dryness and thenresuspended in DCM. The solids were filtered off through a 0.45μ nalgenefilter. The filtrate was concentrated and loaded onto a 24 g RediSepsilica gel column through a 0.45μ nalgene filter. Silica gelchromatography (0%-100% acetone/DCM) afforded the title compound as ayellow foam (0.131 g, 48%).

MS: 304 ES+ (C₃₀H₃₂N₃O₇PS, C₂₁H₂₀P)

Example 10(2S,5R)-2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSodium Sulfate

The Dowex(R) 50WX8-100, ion-exchange resin (25 g, 0.33 mmol) wasconditioned by stirring for 3 hours in 2N sodium hydroxide (61 mL, 0.33mmol). The resin was then loaded into a cartridge and washed with wateruntil the pH was 7. It was then washed with (1/1) acetone/water,followed by water. (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 87, 68 mg, 0.12 mmol) was taken up in water.Acetone was added dropwise until everything was in solution. The yellowsolution was loaded on the resin and washed through with water. Thefractions containing desired product were combined and lyophilized (25mg, 79%) yielding a white solid.

Optical rotation: (0.1 g/dL, MeOH)=−287.

MS: 276 ES− (C₈H₁₀N₃O₆SNa)

¹H NMR (300 MHz, DEUTERIUM OXIDE) δ: 1.76 (dd, J=1.41, 0.85 Hz, 3H)3.40-3.49 (m, 1H) 3.50-3.58 (m, 1H) 4.27 (dd, J=5.09, 2.45 Hz, 1H) 4.44(s, 1H) 6.23-6.31 (m, 1H).

Intermediate 77: (6S)-tert-butyl6-((tert-butyldimethylsilyloxy)methyl)-5-methyl-3-(trimethylsilyloxy)-5,6-dihydropyridine-1(2H)-carboxylate

Methyllithium in Et₂O (73.2 mL, 117.12 mmol) was added dropwise over 20min. to a suspension of copper(I) iodide (11.15 g, 58.56 mmol) in Et₂O(160 mL) and stirred at 0° C. under nitrogen. After 45 min(S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(10 g, 29.28 mmol) in Et₂O (20 mL) was added dropwise and continuedstirring for 45 min. TMs-Cl in THF (58.6 mL, 58.56 mmol) was added,followed by triethylamine (8.16 mL, 58.56 mmol). The resultant mixturewas stirred at rt for 2 h, diluted with ethylacetate washed withice-cold sat. NaHCO₃ (3×) and brine, dried over Na₂SO₄, filtered andconcentrated in vacuo to obtain the desired product as a crude yellowoil (˜12.58 g, 29.27 mmol).

MS: 330 ES+ (C₂₁H₄₃NO₄Si₂)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.05-0.08 (m, 6H) 0.21 (br. s., 9H)0.89 (br. s., 9H) 1.04 (d, J=6.40 Hz, 2H) 1.48 (s, 9H) 2.34 (br. s., 1H)3.42 (br. s., 2H) 3.54 (dd, J=7.44, 3.67 Hz, 1H) 3.91-4.04 (m, 1H)4.07-4.20 (m, 1H) 4.87 (br. s., 1H)

Intermediate 78: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-methyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

The crude (6S)-tert-butyl6-((tert-butyldimethylsilyloxy)methyl)-5-methyl-3-(trimethylsilyloxy)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 77, 12.58 g, 29.27 mmol) in 8 mL ACN was stirred at rtwith Pd(OAc)₂ (6.57 g, 29.27 mmol) for 2 days. The mixture was dilutedwith 160 mL EtOAc, filtered through celite, concentrated in vacuo andsubjected to flash chromatography (220 g, 0-30% EA/Hex) to obtain thedesired product (6.07 g, 58.3%) (over two steps) as a beige solid.

MS: 256 ES+ (C₁₈H₃₃NO₄Si)

1H NMR (300 MHz, CHLOROFORM-d) δ −0.02-0.07 (m, 6H) 0.80-0.91 (m, 9H)1.49 (s, 9H) 2.04 (d, J=1.13 Hz, 3H) 3.69-4.06 (m, 3H) 4.32-4.73 (m, 2H)6.08 (s, 1H)

Intermediate 79: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of cerium(III) chloride heptahydrate (6.36 g,17.07 mmol) and (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-methyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 78, 6.07 g, 17.07 mmol) in MeOH (100 mL) at 0° C., sodiumtetrahydroborate (0.646 g, 17.07 mmol) was added as a solid. The mixturewas stirred at ambient temp for 1 h. The mixture was concentrated anddiluted with NH₄Cl(aq), H₂O and extracted with ether. The ether layerwas separated and washed with brine, dried over Na₂SO₄, filtered andconcentrated to give the desired product (5.48 g, 90%) as a yellow oil.

MS: 258 ES+ (C₁₈H₃₅NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.06 (s, 6H) 0.78-0.94 (m, 9H)1.38-1.50 (m, 9H) 1.56 (br. s., 1H) 1.78 (s, 3H) 2.90-3.39 (m, 1H)3.67-4.28 (m, 5H) 5.80 (br. s., 1H).

Intermediate 80: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred suspension of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 79, 5.48 g, 15.33 mmol),N-(allyloxy)-2-nitrobenzenesulfonamide (7.92 g, 30.65 mmol) andtriphenylphosphine (12.06 g, 45.98 mmol) in toluene (20 mL) was cooledin an ice-bath and added dropwise (E)-diisopropyldiazene-1,2-dicarboxylate (8.91 ml, 45.98 mmol). Reaction was let warmup to rt and continued to stir at rt for 2 h. The reaction mixture wasevaporated and the crude product was loaded onto silica gel, purifiedvia flash chromatography (750 g, 0-50%) to obtain the desired product(7.05 g, 77%) as a yellow oil.

MS: 598 ES+ (C₂₇H₄₃N₃O₈SSi)

Intermediate 81: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 80, 7.05 g, 11.79 mmol) in THF (100 mL) was charged withnitrogen and cooled in an ice-bath. Tetrabutylammonium fluoride in THF(14.15 mL, 14.15 mmol) was added to the solution and stirred at rt. Thereaction mixture was evaporated and the crude product was loaded ontosilica and purified via flash chromatography (30-100% EA/Hexanes, 40 gcolumn), to obtain the desired product (4.52 g, 79%) as a pale yellowfoam.

MS: 484 ES+ (C₂₁H₂₉N₃O₈S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.43 (br. s., 9H) 1.80 (br. s., 3H)3.35 (br. s., 1H) 3.73 (d, J=6.22 Hz, 1H) 3.88 (br. s., 1H) 4.35 (br.s., 5H) 5.15-5.29 (m, 2H) 5.45 (s, 1H) 5.77 (br. s., 1H) 7.61 (d, J=7.35Hz, 1H) 7.76 (dd, J=12.62, 7.54 Hz, 2H) 8.13 (d, J=7.72 Hz, 1H)

Intermediate 82:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-methyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

To a solution of periodic acid (4.10 g, 17.99 mmol) in wet acetonitrile(25 mL) (0.75% water by volume) at room temperature was addedchromium(VI) oxide (0.490 g, 4.90 mmol). The mixture was stirred untilcomplete dissolution was achieved.

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 81, 4.52 g, 9.35 mmol) in wet acetonitrile (25 mL) (0.75%by volume) at 0° C. was added dropwise the previously formed periodicacid/chromium oxide solution. The reaction was stirred o/n at rt. Thereaction mixture was diluted with CHCl₃ and washed with conc. citricacid/water and then with brine (2×). The organics were dried overmagnesium sulfate, filtered and concentrated to obtain the desiredproduct (3.98 g, 86%) as a beige foam.

MS: 498 ES+ (C₂₁H₂₇N₃O₉S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.43 (s, 9H) 1.88 (br. s., 3H) 3.43(d, J=15.07 Hz, 1H) 4.09-4.51 (m, 5H) 4.60-4.92 (m, 1H) 5.14-5.28 (m,2H) 5.46 (br. s., 1H) 5.74 (br. s., 1H) 7.63 (d, J=6.03 Hz, 1H)7.69-7.85 (m, 2H) 8.12 (d, J=7.72 Hz, 1H)

Intermediate 83: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-methyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 82, 3.98 g, 8.00 mmol) in DMF (20 mL) at roomtemperature was added ammonia hydrochloride (0.856 g, 16.00 mmol),2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisouroniumhexafluorophosphate(V) (4.56 g, 12.00 mmol) andN-ethyl-N-isopropylpropan-2-amine (5.57 mL, 32.00 mmol). The reactionmixture was stirred at room temperature for 2 h. The reaction mixturewas diluted with ethyl acetate and washed with saturated sodiumbicarbonate and three times with 1:1 brine:water. The organics weredried over magnesium sulfate, filtered and concentrated. The crudeproduct was purified with silica gel chromatography (220 g, 10%-80%ethyl acetate/hexanes) to yield the desired product (2.79 g, 70.2%) asan yellow foam.

MS: 497 ES+ (C₂₁H₂₈N₄O₈S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.40-1.48 (m, 9H) 1.81 (s, 3H) 3.34(d, J=12.43 Hz, 1H) 4.19-4.53 (m, 4H) 4.84 (br. s., 1H) 5.14-5.38 (m,3H) 5.52 (br. s., 1H) 5.74 (br. s., 1H) 6.45 (br. s., 1H) 7.58-7.64 (m,1H) 7.76 (dtd, J=14.01, 7.69, 7.69, 6.22 Hz, 2H) 8.11 (d, J=7.72 Hz, 1H)

Intermediate 84:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide

(2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-methyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 83, 2.785 g, 5.61 mmol) and zinc(II) bromide (2.53 g,11.22 mmol) was added DCM (10 mL) and stirred at rt overnight. Thereaction mixture was diluted with dichloromethane and washed withsaturated sodium bicarbonate and brine. The organics were dried overmagnesium sulfate, filtered and concentrated to afford the desiredproduct (2.015 g, 91%) as a beige foam.

MS: 397 ES+ (C₁₆H₂₀N₄O₆S)

¹H NMR (300 MHz, CHLOROFORM-d)₆: 1.97 (s, 3H) 2.96 (br. s., 2H) 3.77(br. s., 1H) 4.29 (br. s., 1H) 4.34-4.52 (m, 2H) 5.21-5.44 (m, 4H) 5.55(br. s., 1H) 5.75-5.94 (m, 1H) 7.13 (br. s., 1H) 7.63 (dd, J=7.82, 1.41Hz, 1H) 7.70-7.88 (m, 2H) 8.15 (dd, J=7.72, 1.51 Hz, 1H)

Intermediate 85:(2S,5R)-5-(allyloxyamino)-3-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide

To(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 84, 2.01 g, 5.07 mmol) and potassium carbonate (2.172 g,15.72 mmol) in acetonitrile (10 mL) was added benzenethiol (1.557 mL,15.21 mmol) and stirred at rt for 2 h. The solvent was evaporated at 30°C., redissolved in DCM and a little bit MeOH, filtered and loaded ontosilica gel at 30° C. The crude mixture was purified via flashchromatography (220 g column, 0-20% MeOH/DCM), the fractions wereconcentrated at 35° C. and dried under high-vacuum to obtain the desiredproduct (0.759 g, 70.9%) as an off-white solid.

MS: 212 ES+ (C₁₀H₁₇N₃O₂)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.94 (s, 3H) 2.93-3.09 (m, 2H) 3.40(br. s., 1H) 3.78 (s, 1H) 4.17-4.23 (m, 2H) 5.19-5.25 (m, 1H) 5.25-5.33(m, 1H) 5.37 (br. s., 1H) 5.53 (br. s., 1H) 5.94 (ddt, J=17.00, 10.60,5.98, 5.98 Hz, 1H) 7.14 (br. s., 1H)

Intermediate 86:(2S,5R)-6-(allyloxy)-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

A solution of(2S,5R)-5-(allyloxyamino)-3-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 85, 0.758 g, 3.59 mmol) andN-ethyl-N-isopropylpropan-2-amine (2.507 mL, 14.35 mmol) in acetonitrile(350 mL) was cooled to below 0° C. in an ice-salt bath and added asolution of bis(trichloromethyl) carbonate (0.426 g, 1.44 mmol) in ACN(15 mL) at a rate of 0.1 mL/min. The reaction mixture was stirred at rtovernight. The solvents were evaporated at 30° C., and the crude mixturewas redissolved in EtOAc. The crude organic solution was washed withsat. NaHCO₃, brine, dried over MgSO₄, and filtered. The organics wereevaporated and the crude product was loaded onto silica gel at 30° C.and Purified via flash chromatography (80 g, 0-100% EtOAc/Hexanes), toobtain the desired product (0.700 g, 82%) as pale yellow oil.

MS: 238 ES+ (C₁₁H₁₅N₃O₃)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.92 (d, J=0.75 Hz, 3H) 3.12-3.20 (m,1H) 3.26-3.34 (m, 1H) 3.82 (dd, J=4.99, 2.73 Hz, 1H) 4.30 (s, 1H)4.34-4.50 (m, 2H) 5.28-5.41 (m, 2H) 5.44 (br. s., 1H) 5.94-6.09 (m, 1H)6.09-6.15 (m, 1H) 6.68 (br. s., 1H)

Intermediate 87: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

To a solution of(2S,5R)-6-(allyloxy)-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 86, 200 mg, 0.84 mmol) and AcOH (0.097 mL, 1.69 mmol)(dried over sodium sulfate) in CH₂Cl₂ (7 mL) at room temperature wasadded Pd(Ph₃P)₄ (450 mg, 0.39 mmol). The solution was stirred at rt for45 min. To this reaction mixture was added pyridine (7.00 mL) and sulfurtrioxide pyridine complex (496 mg, 3.12 mmol) and continued stirringunder N₂ at rt overnight. The suspension was evaporated to dryness at39° C. and then resuspended in DCM. The solids were filtered off througha 0.45μ nalgene filter. The filtrate was loaded onto column. Silica gelchromatography (80 g column, 0%-100% acetone/DCM) afforded the desiredproduct (68.0 mg, 13.92%) as an off-white foam.

MS: 276 ES− (C₂₉H₃₀N₃O₆PS)

Example 11(2S,5R)-2-carbamoyl-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSodium Sulfate

(2S,5R)-2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate (4.02 mg, 89%) was prepared a similar manner asdescribed in Example 10 as a white solid, using(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 98, 9 mg, 0.12 mmol).

MS: 304 ES− (C₁₀H₁₄N₃O₆SNa)

¹H NMR (600 MHz, DEUTERIUM OXIDE) δ: 0.95 (d, J=7.15 Hz, 3H) 1.01 (d,J=6.78 Hz, 3H) 2.13 (spt, J=6.78 Hz, 1H) 3.38 (dd, J=11.29, 2.26 Hz, 1H)3.54 (d, J=11.29 Hz, 1H) 4.27 (dd, J=5.27, 2.64 Hz, 1H) 4.49 (s, 1H)6.26 (d, J=5.27 Hz, 1H)

Intermediate 88: (6S)-tert-butyl6-((tert-butyldimethylsilyloxy)methyl)-5-isopropyl-3-(trimethylsilyloxy)-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate (theoretically 6.7 g) was prepared a similar manner asdescribed in Intermediate 77 as a yellow oil, using (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 50, 5 g, 14.64 mmol) as a starting material.

MS: 330 ES+ (C₂₃H₄₇NO₄Si₂)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.05-0.08 (m, 6H) 0.21 (br. s., 9H)0.89 (br. s., 9H) 1.04 (d, J=6.40 Hz, 2H) 1.48 (s, 9H) 2.34 (br. s., 1H)3.42 (br. s., 2H) 3.54 (dd, J=7.44, 3.67 Hz, 1H) 3.91-4.04 (m, 1H)4.07-4.20 (m, 1H) 4.87 (br. s., 1H)

Intermediate 89: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-isopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

(S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-isopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(804 mg, 14.32%) (over two steps) was prepared as described inIntermediate 78 as a yellow oil, using (6S)-tert-butyl6-((tert-butyldimethylsilyloxy)methyl)-5-isopropyl-3-(trimethylsilyloxy)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 88, theoretically 6.7 g, 14.64 mmol).

MS: 384 ES+ (C₂₀H₃₇NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.01 (d, J=3.96 Hz, 6H) 0.84 (s, 9H)1.14-1.23 (m, 6H) 1.49 (s, 9H) 2.35-2.50 (m, 1H) 3.77-3.94 (m, 2H)3.96-4.05 (m, 1H) 4.32-4.85 (m, 2H) 6.05-6.14 (m, 1H)

Intermediate 90: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(0.732 g, 91%), was prepared a similar manner as described forIntermediate 79 as a colorless oil, using (S)-tert-butyl2-((tert-butyldimethylsilyloxy)-methyl)-3-isopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 89, 804 mg, 2.10 mmol) andN-(allyloxy)-2-nitrobenzenesulfonamide (0.979 g, 3.79 mmol) as startingmaterials.

MS: 286 ES+ (C₂₀H₃₉NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.06 (d, J=1.51 Hz, 6H) 0.89 (s, 9H)1.09 (dd, J=10.55, 6.78 Hz, 6H) 1.48 (s, 9H) 2.13-2.26 (m, 1H) 2.86 (br.s., 1H) 3.36-3.52 (m, 1H) 3.60-3.78 (m, 2H) 3.82-4.42 (m, 3H) 5.73-5.91(m, 1H)

Intermediate 91: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(0.928 g, 78%) was prepared as described in Intermediate 80 as a paleyellow oil, using (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 90, 0.731 g, 1.90 mmol) as a starting material.

MS: 626 ES+ (C₂₉H₄₇N₃O₈SSi)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.01 (s, 6H) 0.86 (s, 9H) 0.95-1.09(m, 6H) 1.42 (br. s., 9H) 2.14-2.37 (m, 1H) 3.50 (dd, J=14.60, 4.43 Hz,1H) 3.64-3.89 (m, 2H) 4.21-4.61 (m, 5H) 5.12-5.53 (m, 3H) 5.66-5.88 (m,1H) 7.61 (d, J=7.72 Hz, 1H) 7.68-7.83 (m, 2H) 8.13 (d, J=8.10 Hz, 1H)

Intermediate 92: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(660 mg, 87%) was prepared a similar manner as described in Intermediate81 as a yellow oil, using (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 91, 0.928 g, 1.48 mmol) as a starting material.

MS: 512 ES+ (C₂₃H₃₃N₃O₈S)

Intermediate 93:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

((2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (0.595 g, 87.76%) was prepared a similar manner as described inIntermediate 82 as a yellow oil, using (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 92, 660 mg, 1.29 mmol) as a starting material.

MS: 526 ES+ (C₂₃H₃₁N₃O₉S)

¹H NMR (300 MHz, DMSO-d⁶) δ: 0.62-1.02 (m, 6H) 1.36 (br. s, 9H)2.78-3.26 (m, 1H) 3.34-3.62 (m, 1H) 3.65-4.10 (m, 1H) 4.10-4.45 (m, 3H)4.46-4.84 (m, 1H) 5.01-5.64 (m, 3H) 5.64-5.99 (m, 1H) 7.87-8.14 (m, 4H)13.03 (br. s, 1H)

Intermediate 94: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(0.260 g, 43.7%) was prepared a similar manner as described inIntermediate 83 as a pale yellow oil, using(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 93, 0.595 g, 1.13 mmol) as a starting material.

MS: 525 ES+ (C₂₃H₃₂N₄O₈S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.02 (d, J=6.97 Hz, 6H) 1.44 (br. s.,9H) 2.24 (s, 1H) 3.29-3.40 (m, 1H) 4.18-4.50 (m, 4H) 5.02 (br. s., 1H)5.15-5.28 (m, 3H) 5.73 (s, 2H) 6.44 (s, 1H) 7.60-7.64 (m, 1H) 7.76 (dqd,J=14.76, 7.54, 7.54, 7.54, 1.60 Hz, 2H) 8.11 (dd, J=7.91, 1.32 Hz, 1H)

Intermediate 95:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(160 mg, 86%) was prepared a similar manner as described in Intermediate84 as a beige solid, using (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-isopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 94, 260 mg, 0.5 mmol) as a starting material.

MS: 425 ES+ (C₁₈H₂₄N₄O₆S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.01-1.12 (m, 6H) 2.64-2.77 (m, 1H)2.82-3.06 (m, 2H) 3.93 (s, 1H) 4.33 (br. s., 1H) 4.35-4.55 (m, 2H)5.21-5.31 (m, 3H) 5.59 (br. s., 1H) 5.76-5.93 (m, 1H) 7.04 (br. s., 1H)7.62 (dd, J=7.54, 1.51 Hz, 1H) 7.71-7.85 (m, 2H) 8.14 (dd, J=7.82, 1.41Hz, 1H)

Intermediate 96:(2S,5R)-5-(allyloxyamino)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

(2S,5R)-5-(allyloxyamino)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(55.0 mg, 61.0%) was prepared a similar manner as described inIntermediate 85 as an off-white solid, using(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 95, 160 mg, 0.38 mmol) as a starting material.

MS: 340 ES+ (C₁₂H₂₁N₃O₂)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.05 (d, J=6.97 Hz, 3H) 1.11 (d,J=6.78 Hz, 3H) 2.54-2.68 (m, 1H) 2.94-3.11 (m, 2H) 3.45 (br. s, 1H) 3.95(br. s., 1H) 4.20 (d, J=5.84 Hz, 2H) 5.22 (d, J=10.36 Hz, 1H) 5.29 (dd,J=17.14, 1.51 Hz, 1H) 5.38 (br. s., 1H) 5.58 (d, J=3.58 Hz, 1H)5.87-6.02 (m, 1H) 7.00 (br. s., 1H)

Intermediate 97:(2S,5R)-6-(allyloxy)-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

(2S,5R)-6-(allyloxy)-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(0.46 mg, 77%) was prepared a similar manner as described inIntermediate 86 as a pale yellow oil, using(2S,5R)-5-(allyloxyamino)-3-isopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 96, 0.54 mg, 0.23 mmol) as a starting material.

MS: 266 ES+ (C₁₃H₁₉N₃O₃)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.02 (d, J=6.78 Hz, 3H) 1.10 (d,J=6.78 Hz, 3H) 2.60 (quin, J=6.69 Hz, 1H) 3.14-3.22 (m, 1H) 3.24-3.32(m, 1H) 3.86 (dd, J=5.27, 2.83 Hz, 1H) 4.33-4.50 (m, 3H) 5.26-5.52 (m,3H) 5.94-6.15 (m, 2H) 6.63 (br. s., 1H)

Intermediate 98: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (9.00 mg, 8.73%) was prepared in a similar manner as describedin Intermediate 87 as a colorless oil, using(2S,5R)-6-(allyloxy)-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 97, 45 mg, 0.17 mmol) as a starting material.

MS: 304 ES− (C₃₁H₃₄N₃O₆PS)

Example 12(2S,5R)-4-carbamoyl-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate, Monosodium Salt

A stirred solution of(2S,5R)-6-hydroxy-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide(Intermediate 116, 21 mg, 0.09 mmol) in pyridine (2 mL) was prepared andplaced under a nitrogen atmosphere. To it was added Sulfurtrioxide-Pyridine complex (6 eq, 88 mg, 0.55 mmol). This mixture wasstirred overnight at ambient temperature; the mixture was thenconcentrated under reduced pressure and the residue dissolved in a smallvolume of water. A Dowex column was prepared as follows: Dowex(R)50WX8-100, ion-exchange resin (16 g) was conditioned by stirring for 3hours in 2N sodium hydroxide (34 mL). The resin was then loaded into acartridge and washed with water until pH 7. It was then washed with(1/1) acetone/water, followed by water again. The aqueous solution ofcrude product was applied to the top of this column; the desired productwas eluted with water. The fractions containing desired product werecombined and lyophilized. The obtained material was triturated withmethanol and the filtrate concentrated and freeze-dried again. Thedesired product was obtained as a white solid (19 mg, 62%).

MS: 308 ES+ (C₉H₁₃N₃O₇S)

¹H NMR (300 MHz, D₂O) δ: 3.37-3.50 (m, 4H) 3.51-3.63 (m, 2H) 3.72-3.88(m, 3H) 4.26 (ddd, J=7.79, 4.67, 3.21 Hz, 1H) 4.77 (d, J=1.51 Hz, 1H)6.61 (d, J=2.83 Hz, 1H)

Intermediate 99: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-oxoethyl)carbamate

A stirred solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 5, 28.36 g, 70.44 mmol) in methylene chloride (247 mL) wasprepared, cooled in a dry ice/acetone bath, and placed under nitrogen.To it was added diisobutylaluminum hydride (1.0 M in methylene chloride)(106 mL, 105.66 mmol); the mixture was maintained at that temperaturefor about 2.5 hours. The reaction was monitored by TLC (25% ethylacetate in hexanes). Upon complete conversion of starting material, thereaction was quenched with methanol. The resulting mixture was dilutedwith methylene chloride (about 400 mL) and washed with a 10% aqueoussolution (w/w) of Rochelle's salt. The organic layer was separated andthe aqueous layer was back-extracted with methylene chloride (about 250mL). The two organic layers were combined, washed with brine, dried oversodium sulfate, filtered, and concentrated in vacuo. The desired productwas isolated using normal-phase chromatography (330 g column, 45minutes, 5-45% ethyl acetate in hexanes). The obtained product waspurified a second time by normal-phase chromatography (0-20% acetone inhexanes, 120-g column), affording a yellow oil, 16.1 g (66%).

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H) 0.84 (s, 9H) 1.30-1.46 (m, 9H)3.65-3.91 (m, 4H) 4.43-4.73 (m, 1H) 5.17 (dt, J=8.12, 1.61 Hz, 1H) 5.21(dd, J=1.70, 0.76 Hz, 1H) 5.69-5.87 (m, 1H) 9.38-9.50 (m, 1H)

Intermediate 100: methyl4-(tert-butoxycarbonyl((S)-1-(tert-butyldimethylsilyloxy)but-3-en-2-yl)amino)-3-hydroxy-2-methylenebutanoate

To a stirred solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-oxoethyl)carbamate(Intermediate 99, 16.1 g, 46.8 mmol), methyl acrylate (6.3 mL, 70 mmol),and methanol (about 2 mL), under nitrogen at ambient temperature, wasadded Quinuclidine (2.1 g, 19 mmol) as a solid. The mixture was thenstirred for 1 day; analysis by TLC (15% ethyl acetate in hexanes)indicated incomplete conversion of starting material. Another 0.8 eq ofquinuclidine was added every 24 hours; another 0.75 eq methyl acrylateand 1 mL methanol were added every two days. After 6 days total, thereaction was diluted with water and extracted with ethyl acetate. TLCindicated that only one extraction with about 250 mL ethyl acetate wasneeded to bring all the crude product into the organic layer, which wasthen washed with brine, dried over sodium sulfate, and concentratedunder reduced pressure. Normal-phase chromatography (5-30% ethyl acetatein hexanes, 220-g column, 35 minutes) afforded the desired compound (16g, 80%, pale oil) as a mixture of diastereomers, with about 10 mol % ofstarting aldehyde present according to proton NMR analysis. The materialwas taken forward in this state.

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H) 0.85 (s, 9H) 1.37 (s, 9H)3.19-3.47 (m, 2H) 3.68 (s, 3H) 3.73-3.92 (m, 2H) 3.95-4.19 (m, 1H)4.47-4.61 (m, 1H) 5.03-5.25 (m, 3H) 5.78-5.95 (m, 2H) 6.16 (br. s, 1H)

Intermediate 101: (2S,5S)-1-tert-butyl 4-methyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1,4(2H)-dicarboxylate

A stirred solution of methyl4-(tert-butoxycarbonyl((S)-1-(tert-butyldimethylsilyloxy)but-3-en-2-yl)amino)-3-hydroxy-2-methylenebutanoate(Intermediate 100, 16 g, 37 mmol) in toluene (about 300 mL), at ambienttemperature in a heat-dried flask, was thoroughly degassed with argonand placed under a nitrogen atmosphere. To this was added Hoveyda-GrubbsCatalyst 2nd Generation (460 mg, 0.74 mmol). Analysis of the mixture 3.5hours later by LCMS and TLC (15% ethyl acetate in hexanes) indicatedvery high conversion of starting material to two major products. Thereaction mixture was concentrated in vacuo in the presence of enoughsilica gel to give a free-flowing powder upon reaching dryness.Normal-phase chromatography (0-100% ethyl acetate in hexanes, 330-gcolumn, 45 minutes) was used to isolate the two compounds; thediastereomer shown above (6.4 g, 43%, dark oil) eluted first.

MS: 402 ES+ (C₁₉H₃₅NO₆Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (s, 6H) 0.83 (s, 9H) 1.41 (s, 9H)2.81-3.10 (m, 1H) 3.68-3.78 (m, 5H) 3.93-4.15 (m, 1H) 4.34 (br. s., 1H)4.42-4.64 (m, 1H) 4.99 (d, J=5.48 Hz, 1H) 6.94 (d, J=6.80 Hz, 1H)

Intermediate 102: (2S,5R)-1-tert-butyl 4-methyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1,4(2H)-dicarboxylate

This product (6.6 g, 44%, dark oil) was prepared in the same reaction asIntermediate 101, and eluted second in the chromatographic purificationdescribed in that procedure.

MS: 402 ES+ (C₁₉H₃₅NO₆Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.04 (s, 6H) 0.86 (s, 9H) 1.40 (s, 9H) 2.80(br. s., 1H) 3.68 (s, 3H) 3.76 (d, J=5.67 Hz, 2H) 4.03 (m, J=7.20 Hz,1H) 4.20-4.33 (m, 1H) 4.42 (br. s., 1H) 5.24 (d, J=5.85 Hz, 1H) 6.64(br. d, J=1.00 Hz, 1H)

Intermediate 103:(3S,6S)-1-(tert-butoxycarbonyl)-6-((tert-butyldimethylsilyloxy)methyl)-3-hydroxy-1,2,3,6-tetrahydropyridine-4-carboxylicacid

A stirred solution of (2S,5S)-1-tert-butyl 4-methyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1,4(2H)-dicarboxylate(Intermediate 101, 5.9 g, 15 mmol) in THF (55 mL) and water (18 mL) wasprepared under ambient conditions. To it was added lithium hydroxide(700 mg, 29 mmol) as a solid. The mixture was stirred under anatmosphere of nitrogen at ambient temperature. The solid materialdissolved within a few minutes. Analysis of the mixture by TLC (25%ethyl acetate in hexanes) at 30 minutes indicated complete consumptionof starting material. About 20 mL of a 1N aq solution of HCl was addedto the mixture as it stirred. A 10% aq solution of citric acid was usedto bring the pH to the 4-5 range. The mixture was diluted with about 60mL water and extracted twice with ethyl acetate. The organic extractswere combined, washed with brine, dried over sodium sulfate, filtered,and concentrated in vacuo. The dark brown, gummy residue (5.5 g, 96%,dark oil) was characterized by proton NMR and LCMS as desired product,and taken forward without further purification.

MS: 386 ES− (C₁₈H₃₃NO₆Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.04 (s, 6H) 0.86 (s, 9H) 1.40 (s, 9H) 2.82(br. s., 1H) 3.30 (br. s., 2H) 3.75 (d, J=6.04 Hz, 2H) 3.96-4.11 (m, 1H)4.25 (t, J=7.55 Hz, 1H) 4.41 (br. s., 1H) 6.65 (d, J=3.59 Hz, 1H)

Intermediate 104: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-carbamoyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

A stirred solution of(3S,6S)-1-(tert-butoxycarbonyl)-6-((tert-butyldimethylsilyloxy)methyl)-3-hydroxy-1,2,3,6-tetrahydropyridine-4-carboxylicacid (Intermediate 103, 5.5 g, 14 mmol) and N,N-diisopropylethylamine(7.4 mL, 42 mmol) in DMF (about 50 mL) was prepared under ambientconditions. To this was addedO-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (8.0 g, 21 mmol). The reaction vessel was purgedwith argon for one minute, and the dark, clear mixture was stirred undernitrogen for 5 minutes at ambient temperature. To the mixture was thenadded ammonium chloride (1.5 g, 22 mmol). The mixture was stirred undernitrogen at ambient temperature overnight. In the morning, analysis byLCMS indicated consumption of starting material and the presence of onemajor product with a longer retention time and desired mass. Aqueousworkup using diethyl ether separated crude product from water-solublematerials. The organic phases were combined, washed with brine, driedover sodium sulfate, filtered, and concentrated in vacuo. Normal-phasechromatography (1-6% methanol in methylene chloride, 220-g, about 50min) afforded the desired product (2.4 g, 44%, tan solid).

MS: 387 ES+ (C₁₈H₃₄N₂O₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.04 (s, 6H) 0.86 (s, 9H) 1.40 (s, 9H) 2.74(br. s., 1H) 3.72 (d, J=5.48 Hz, 2H) 4.09 (br. s., 1H) 4.18-4.32 (m, 1H)4.40 (br. s., 1H) 5.59 (d, J=5.48 Hz, 1H) 6.54 (d, J=3.02 Hz, 1H) 7.15(br. s., 1H) 7.36 (br. s., 1H)

Intermediate 105: (2S,5R)-tert-butyl5-(N-(allyloxy)-2,4-dinitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate

A solution of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-carbamoyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 104, 1.4 g, 3.6 mmol) in toluene (35 mL) was prepared andstirred under nitrogen at ambient temperature. To it was addedtriphenylphosphine (1.1 g, 4.3 mmol) andN-(allyloxy)-2,4-dinitrobenzenesulfonamide (1.1 g, 3.6 mmol). Upondissolution of these materials, the mixture was stirred for another tenminutes. To the mixture was then added diisopropyl azodicarboxylate (840μl, 4.3 mmol) by syringe. Argon was blown over the top of the mixtureand the mixture was then placed under nitrogen and stirred overnight.Analysis by LCMS in the morning and also by TLC (5% methanol inmethylene chloride) indicated consumption of starting alcohol. Thereaction mixture was adsorbed onto silica gel and normal phasechromatography (15-75% ethyl acetate in hexanes, 220-g column) affordedthe desired product (orange solid; ˜100% yield).

MS: 673 ES+ (C₂₇H₄₁N₅O₁₁SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (s, 6H) 0.84 (s, 9H) 1.29-1.49 (m, 9H)3.62-3.78 (m, 2H) 4.00 (s, 1H) 4.15-4.87 (m, 4H) 5.16-5.36 (m, 2H)5.67-5.90 (m, 1H) 6.76-6.88 (m, 1H) 6.99-7.80 (m, 2H) 8.24-8.38 (m, 1H)8.62 (ddd, J=8.64, 6.18, 2.36 Hz, 1H) 9.04 (d, J=2.27 Hz, 1H)

Intermediate 106:(3R,6S)-3-(N-(allyloxy)-2,4-dinitrophenylsulfonamido)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridine-4-carboxamide

A stirred solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2,4-dinitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 105, 1.8 g, 2.6 mmol) in methylene chloride (about 25 mL)was prepared under ambient conditions; argon was blown over the solutionfor 30 seconds, and the solution was then stirred at ambienttemperature. To it was added zinc bromide (4.1 g, 18 mmol). The mixturewas stirred overnight. In the morning, solid, gummy, yellow materialcoated the bottom of the flask, and analysis by LCMS indicatedincomplete conversion. A small amount of THF (about 2 mL) was added. Themixture became homogenous over the next 25 seconds. Analysis over thenext few hours indicated increased reaction rate with little or noincrease in by-product formation. More zinc bromide (another 5 g) wasadded in parts over that time. If addition of zinc bromide was followedby precipitation, another small volume of THF was added. By the end ofthe day, consumption of starting material was still incomplete but high.The reaction was carefully transferred into a saturated solution ofsodium bicarbonate. A great deal of gas evolution was observed. Crudeproduct was extracted using ethyl acetate until TLC (5% methanol inmethylene chloride) indicated no UV-active material remained in theaqueous. The organic extracts were combined, washed with brine, driedover sodium sulfate, filtered, and concentrated. The residue wasdissolved in methylene chloride for normal-phase chromatography (0-4%methanol in methylene chloride over 35 min, 80-g column), affordingdesired compound (1.1 g, 72%, pale yellow solid).

MS: 573 ES+ (C₂₂H₃₃N₅O₉SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.01-0.10 (m, 6H) 0.78-0.91 (m, 9H) 2.85(br. s., 2H) 3.38-3.66 (m, 3H) 4.31-4.49 (m, 2H) 4.60 (br. s., 1H)5.17-5.39 (m, 3H) 5.78-5.97 (m, 1H) 6.73-6.91 (m, 2H) 7.36 (br. s., 1H)8.32 (d, J=8.69 Hz, 1H) 8.62 (dd, J=8.88, 2.27 Hz, 1H) 9.01 (d, J=2.27Hz, 1H)

Intermediate 107:(3R,6S)-3-(allyloxyamino)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridine-4-carboxamide

To a reddish-brown, stirred suspension of(3R,6S)-3-(N-(allyloxy)-2,4-dinitrophenylsulfonamido)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridine-4-carboxamide(Intermediate 106, 1.1 g, 1.9 mmol) and cesium carbonate (3.1 g, 9.5mmol) in THF (about 45 mL), at ambient temperature under a blanket ofargon, was added benzenethiol (polymer-bound reagent, 1.55 mmol/g) (4.5g, 7.0 mmol). This mixture was stirred overnight at ambient temperature.Analysis by LCMS in the morning indicated complete consumption ofstarting material, as did analysis by TLC (5% methanol in methylenechloride). The mixture was filtered and the filtered material was rinsedwith THF until no material remained that was responsive to UV or I₂stain on a silica plate. The red-orange solution obtained wasconcentrated and redissolved in methylene chloride. Normal-phasechromatography (0-6% methanol in methylene chloride, 80-g, 25 min.)afforded the desired compound (510 mg, 80%, orange-brown solid).

MS: 343 ES+ (C₁₆H₃₁N₃O₃Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.04 (s, 6H) 0.87 (s, 9H) 2.39 (br. s., 1H)2.74 (dd, J=12.84, 3.40 Hz, 1H) 2.94 (dd, J=12.84, 3.40 Hz, 1H)3.31-3.37 (m, 1H) 3.49-3.57 (m, 2H) 3.57-3.66 (m, 1H) 4.11 (dt, J=5.62,1.16 Hz, 2H) 5.13 (ddt, J=10.50, 2.10, 1.16, 1.16 Hz, 1H) 5.22 (dq,J=17.37, 1.70 Hz, 1H) 5.92 (ddt, J=17.37, 10.39, 5.67, 5.67 Hz, 1H) 6.41(d, J=8.50 Hz, 1H) 6.68 (dd, J=3.21, 0.57 Hz, 1H) 6.98 (br. s., 1H) 7.36(br. s., 1H)

Intermediate 108:(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide

A stirred solution of(3R,6S)-3-(allyloxyamino)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridine-4-carboxamide(Intermediate 107, 505 mg, 1.48 mmol) and N,N-diisopropylethylamine(1.03 mL, 5.91 mmol) in acetonitrile (150 mL) was prepared. Over thiswas blown argon for one minute. The solution was then placed under anargon atmosphere and cooled in an ice bath for 15 minutes. To it wasadded a solution of triphosgene (180 mg, 0.59 mmol) in acetonitrile (10mL) via syringe using a syringe pump set to deliver 0.1 mL/min. Duringthe course of the addition, temperature was kept at or near 0° C. Uponaddition of all triphosgene, the orange solution was stirred at 0° C.for another 30 minutes. The ice bath was then removed and the mixturewas stirred for another 30 minutes at ambient temperature. Analysis atthat time by LCMS indicated conversion to desired product. The reactionmixture was concentrated in vacuo and the residue was dissolved inmethylene chloride. Normal-phase chromatography (15-65% ethyl acetate inmethylene chloride, 40-g column, 25 minutes) was used to isolate thedesired product (454 mg, 84%, white solid).

MS: 349 ES+ (C₁₇H₂₉N₃O₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.06 (s, 6H) 0.87 (s, 9H) 3.11-3.20 (m, 1H)3.25-3.35 (m, 1H) 3.71-3.79 (m, 1H) 3.79-3.95 (m, 2H) 4.34 (dt, J=5.90,1.20 Hz, 2H) 4.51 (d, J=2.83 Hz, 1H) 5.23 (ddt, J=10.43, 1.89, 1.01,1.01 Hz, 1H) 5.33 (dq, J=17.30, 1.60 Hz, 1H) 5.80-6.00 (m, 1H) 6.49 (dd,J=2.83, 0.94 Hz, 1H) 7.08 (br. s., 1H) 7.51 (br. s., 1H)

Intermediate 109:(2S,5R)-6-(allyloxy)-2-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide

(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide(Intermediate 108, 450 mg, 1.2 mmol) was dissolved in THF (about 7 mL);the atmosphere in the reaction vessel was evacuated and backfilled withargon, and the solution was stirred in an ice bath for ten minutes. Toit was then added tetrabutylammonium fluoride (1 M in THF) (1.5 mL, 1.5mmol) dropwise. After one hour, analysis of the mixture by LCMSindicated consumption of starting material and formation of one majorproduct. The mixture was concentrated and dissolved in methylenechloride. Normal-phase chromatography (0-10% methanol in methylenechloride, 25-g column, 25 minutes) was used to isolate the desiredproduct (284 mg, 92%, white solid).

MS: 254 ES+ (C₁₁H₁₅N₃O₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.04-3.28 (m, 2H) 3.54-3.80 (m, 3H) 4.34(dt, J=6.00, 1.16 Hz, 2H) 4.50 (d, J=2.83 Hz, 1H) 5.01-5.10 (m, 1H)5.18-5.27 (m, 1H) 5.33 (dq, J=17.28, 1.54 Hz, 1H) 5.78-6.02 (m, 1H) 6.52(d, J=1.70 Hz, 1H) 7.06 (br. s., 1H) 7.49 (br. s., 1H)

Intermediate 110: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound (4.0 g, 74%) was prepared from (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-carbamoyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 104, 3.36 g, 8.69 mmol) using the same procedure toprepare Intermediate 105 from Intermediate 104, but usingN-(allyloxy)-2-nitrobenzenesulfonamide (Intermediate 9, 2.24 g, 8.69mmol) as a reagent in place ofN-(allyloxy)-2,4-dinitrobenzenesulfonamide.

MS: 627 ES+ (C₂₇H₄₂N₄O₉SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (s, 6H) 0.84 (s, 9H) 1.36 (br. s., 9H)2.84-3.22 (m, 1H) 3.59-3.77 (m, 2H) 4.10-4.46 (m, 3H) 4.64 (br. s., 1H)4.82 (d, J=13.22 Hz, 1H) 5.08-5.42 (m, 2H) 5.63-5.93 (m, 1H) 6.72-6.83(m, 1H) 7.37 (br. s., 1H) 7.49-7.71 (m, 1H) 7.85 (dt, J=8.21, 4.01 Hz,1H) 7.97 (br. s., 2H) 8.00-8.11 (m, 1H)

Intermediate 111: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-carbamoyl-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound (617 mg, 67%) was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 110, 1.2 g, 1.80 mmol) using the same procedure to prepareIntermediate 109 from Intermediate 108.

MS: 513 ES+ (C₂₁H₂₈N₄O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.19-1.56 (m, 9H) 2.86-3.20 (m, 1H)3.44-3.55 (m, 2H) 3.89-4.65 (m, 4H) 4.82 (d, J=11.14 Hz, 1H) 4.94-5.07(m, 1H) 5.12-5.36 (m, 2H) 5.57-5.92 (m, 1H) 6.70-7.10 (m, 2H) 7.36 (br.s., 1H) 7.78-7.89 (m, 1H) 7.96 (d, J=2.83 Hz, 2H) 8.00-8.14 (m, 1H)

Intermediate 112: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-carbamoyl-2-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

A solution of Intermediate 111 (427 mg, 0.83 mmol) and methyl iodide (6eq) (311 μA, 5.00 mmol) in acetonitrile (˜4 mL) was prepared underambient conditions and placed under a nitrogen atmosphere. In thismixture was suspended silver oxide (1.1 eq) (212 mg, 0.92 mmol). Themixture was protected from light through the use of aluminum foil andstirred until analysis by TLC (1:1 acetonitrile:DCM) and LCMS indicatedcomplete consumption of starting material, around 3 days. Filtration ofthe reaction mixture through a 0.45μ filter gave a bronze solution. Thiswas concentrated in vacuo and redissolved in methylene chloride. Thecrude mixture was purified by normal-phase chromatography (10-50%acetonitrile in DCM) to afford the desired product as a colorlessresidue (109 mg, 25%). The isolated material was lyophilized (whitepowder) for ease of characterization.

MS: 527 ES+ (C₂₂H₃₀N₄O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.38 (d, J=6.99 Hz, 9H) 2.82-3.15 (m, 1H)3.26 (s, 3H) 3.44 (d, J=3.02 Hz, 2H) 3.90-4.39 (m, 3H) 4.46-4.94 (m, 2H)5.13-5.33 (m, 2H) 5.64-5.89 (m, 1H) 6.71-6.84 (m, 1H) 6.86-7.15 (m, 1H)7.35 (br. s., 1H) 7.85 (t, J=8.03 Hz, 1H) 7.90-8.15 (m, 3H)

Intermediate 113:(3R,6S)-3-(N-(allyloxy)-2-nitrophenylsulfonamido)-6-(methoxymethyl)-1,2,3,6-tetrahydropyridine-4-carboxamide

To a solution of Intermediate 112 (729 mg, 1.38 mmol) in dichloromethane(10 mL) was added trifluoroacetic acid (2133 μl, 27.69 mmol). Thissolution was slightly darker after addition. The reaction was placedunder a nitrogen atmosphere and stirred until LCMS analysis indicatedconsumption of starting material (˜45 minutes). At that time, solventwas removed in vacuo. The residue was taken up in toluene and sonicated,and then toluene was removed in vacuo. This was repeated twice,affording an orange solid of high purity by proton NMR. The product wastaken forward without further purification.

MS: 427 ES+ (C₁₇H₂₂N₄O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.24-3.38 (m, 5H) 3.53-3.71 (m, 2H) 4.28(m, J=11.10, 6.40 Hz, 2H) 4.34-4.44 (m, 1H) 5.11 (br. s., 1H) 5.20-5.35(m, 2H) 5.84 (ddt, J=17.14, 10.43, 6.42, 6.42 Hz, 1H) 6.61 (d, J=1.70Hz, 1H) 7.28 (br. s., 1H) 7.65 (br. s., 1H) 7.88-7.99 (m, 1H) 8.01-8.15(m, 3H) 8.85-9.45 (m, 1H)

Intermediate 114:(3R,6S)-3-(allyloxyamino)-6-(methoxymethyl)-1,2,3,6-tetrahydro-pyridine-4-carboxamide

This intermediate was prepared from(3R,6S)-3-(N-(allyloxy)-2-nitrophenylsulfonamido)-6-(methoxymethyl)-1,2,3,6-tetrahydropyridine-4-carboxamide(Intermediate 113, 734 mg, 1.36 mmol) using the same procedure andisolation techniques to prepare Intermediate 107 from Intermediate 106.The title compound was an orange oil (173 mg, 53%).

MS: 242 ES+ (C₁₁H₁₉N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.70-2.79 (m, 1H) 2.70-2.78 (m, 1H)2.89-2.99 (m, 1H) 2.94 (dd, J=12.94, 3.30 Hz, 1H) 3.22-3.53 (m, 4H) 3.26(s, 3H) 3.61 (br. s., 1H) 4.11 (dt, J=5.62, 1.25 Hz, 2H) 5.09-5.29 (m,2H) 5.92 (ddt, J=17.33, 10.48, 5.74, 5.74 Hz, 1H) 6.34-6.49 (m, 1H) 6.65(dd, J=3.21, 0.76 Hz, 1H) 7.00 (br. s., 1H) 7.39 (br. s., 1H)

Intermediate 115:(2S,5R)-6-(allyloxy)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide

A solution of(3R,6S)-3-(allyloxyamino)-6-(methoxymethyl)-1,2,3,6-tetrahydro-pyridine-4-carboxamide(Intermediate 114, 125 mg, 0.52 mmol) and triethylamine (210 mg, 2.07mmol) in acetonitrile (45 mL) was prepared. Over this was blown argonfor one minute. The solution was then placed under an argon atmosphereand cooled in an ice bath for 15 minutes. To it was slowly added asolution of diphosgene (56 mg, 0.28 mmol) in 5 mL acetonitrile viasyringe pump at a rate of 0.1 mL/min. Once the addition was complete,the reaction was stirred at room temperature overnight. The reactionmixture was then concentrated in vacuo. The residue was dissolved inmethylene chloride and subjected to normal-phase chromatography (0-7%methanol in DCM) to afford 60 mg (43%) of a pale yellow foam uponapplication of high vacuum.

MS: 268 ES+ (C₁₂H₁₇N₃O₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 3.27-3.42 (m, 5H) 3.59-3.77 (m, 2H)4.07-4.15 (m, 1H) 4.36-4.55 (m, 3H) 5.27-5.31 (m, 1H) 5.32-5.36 (m, 1H)5.38-5.70 (m, 2H) 5.92-6.13 (m, 1H) 6.39 (dd, J=3.02, 1.13 Hz, 1H)

Intermediate 116:(2S,5R)-6-hydroxy-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide

A stirred solution of Intermediate 115 (60 mg, 0.22 mmol) indichloromethane (2 mL) was prepared and placed under a nitrogenatmosphere. To it was added acetic acid (25.7 μl, 0.45 mmol) followed bytetrakis(triphenylphosphine)palladium(0) (259 mg, 0.22 mmol). Themixture was stirred under nitrogen at ambient temperature for about 60minutes. Upon complete consumption of starting material as indicated byLCMS analysis, the solvent was removed in vacuo. Addition of MeCNprecipitated a tan solid, which was filtered and discarded. The filtratewas adsorbed onto Celite, and the desired product (21 mg, 41%, whitesolid) was isolated using reverse-phase chromatography (100% water)followed by freeze-drying.

MS: 228 ES+ (C₉H₁₃N₃O₄)

¹H NMR (300 MHz, METHANOL-d₄) δ: 3.38 (dt, J=3.26, 1.68 Hz, 2H) 3.46 (s,3H) 3.68-3.83 (m, 2H) 4.05 (td, J=5.76, 3.02 Hz, 1H) 4.38-4.44 (m, 1H)6.58 (dd, J=2.93, 1.04 Hz, 1H)

Example 13(2S,5R)-2,4-bis(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate Sodium Salt

To a solution of Intermediate 124 (100 mg, 0.37 mmol) in DCM (3 mL) wasadded acetic acid (0.043 mL, 0.75 mmol) (dried over Na₂SO₄) andPd(PPh₃)₄ (215 mg, 0.19 mmol) under argon at room temperature. Theyellow solution was stirred at room temperature for 1 hr. TLC and LCMSshowed complete conversion of starting material to product. The reactionmixture was concentrated and dried under vacuum. Then it was dilutedwith 5 mL 10% CH₃CN in water and loaded onto 30 g ISCO gold C18 column,and eluted with 0% CH₃CN for 5 minutes, followed by 0-50% CH₃CN/H₂O over15 minutes. The first few fractions were collected and lyophilized togive an yellow oil (126 mg). The oil was dissolved in pyridine (3 mL) atrt. Then SO₃-Pyr (353 mg, 2.22 mmol) was added as solid. The mixture wasthen stirred at rt overnight. LCMS showed the completion of thereaction. The reaction solution was concentrated to dryness andazeotropic with 2×5 mL toluene. The crude was then applied to ˜30 gpretreated dowex resin (200 mL 2N NaOH for 2 hrs, then packed and washedwith H₂O til neutral pH). And purified using a gravity column with H₂O,collecting every 10 mL. Fractions containing the desired compound werecombined and lyophilized to give a white solid, which was then purifiedby RP-HPLC to give ˜20 mg white solid.

MS: 309 ES+ (C₁₀H₁₆N₂O₇S)

¹H NMR (300 MHz, D₂O) δ: 3.31 (s, 3H) 3.40 (s, 3H) 3.44 (s, 2H)3.57-3.77 (m, 2H) 3.96-4.12 (m, 3H) 4.24 (s, 1H) 5.62 (br. s., 1H)

Intermediate 117: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(hydroxymethyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 7, 18 g, 52.71 mmol) in a mixture of DCM (900 mL) and MeOH(350 mL) was added successively formaldehyde (42.8 mL, 527.06 mmol) andtributylphosphine (0.692 mL, 3.16 mmol) at rt. The solution was stirredat room temperature for 2-3 hr until TLC (4:1 Hex/EA) showed that thereaction was complete. It was concentrated under reduced pressure andpurified by 120 g silica gel column (0-50% Hex/EA) to afford the titlecompound (20.00 g, 102%) as an oil.

MS: 372 ES+ (C₁₈H₃₃NO₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.13-0.08 (m, 6H) 0.73-0.97 (m, 9H) 1.43(s, 9H) 3.87 (m, 3H) 4.11 (m, 2H) 4.22-4.39 (m, 1H) 4.69-4.86 (m, 1H)5.01 (m, 1H) 6.92-7.07 (m, 1 H).

Intermediate 118: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of Intermediate 117 (7.3 g, 19.65 mmol) in DCM(180 mL) was added N1,N1,N8,N8-tetramethylnaphthalene-1,8-diamine (25.3g, 117.89 mmol). Then it was cooled to 0° C. and trimethyloxoniumtetrafluoroborate (8.72 g, 58.94 mmol) was added. The reaction was thenstirred at rt for several hours until LCMS showed the starting materialwas all consumed. Then it was concentrated under reduced pressure. Theresidue was then taken up in 100 mL Et₂O and filtered, washed with 100mL Et₂O. The organic layer was then washed with 10% citric acid, aqNaHCO₃, brine, dried over MgSO₄, filtered and concentrated. The residuewas then taken up in MeOH (180 mL), cooled to 0° C. and cerium(III)chloride (7.32 g, 19.65 mmol) was then added to give a clear solution.Then NaBH₄ (0.743 g, 19.65 mmol) was added as solid, and the mixture wasstirred from 0° C. to rt for 1 hr. The reaction mixture was concentratedin vacuo. The white solid was redissolved in 200 mL Et₂O and washed with10% citric acid, NaHCO₃, brine, dried over MgSO₄, filtered andconcentrated. The residue was purified by silica gel column (20-80%Hex/EA, 80 g) to afford the title compound (6.20 g, 81%) as a colorlessoil (−3.8:1 mixture of 2 diastereomers).

MS: 388 ES+ (C₁₉H₃₇NO₅Si)

Intermediate 119: (S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of Intermediate 118 (6.2 g, 16.00 mmol),N-(allyloxy)-2-nitrobenzenesulfonamide (4.34 g, 16.80 mmol) andtriphenylphosphine (5.03 g, 19.20 mmol) in toluene (140 mL), was added(E)-diisopropyl diazene-1,2-dicarboxylate (3.67 g, 17.60 mmol) at rt.The reaction was stirred at rt over the weekend. The mixture was thenconcentrated and the residue was purified by silica gel column (120 g,0-100% Hex/EA) to give the title compound (8.80 g, 88%) as an oil

MS: 628 ES+ (C₂₈H₄₅N₃O₉SSi)

Intermediate 120:(S)—N-(allyloxy)-N-(6-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-1,2,3,6-tetrahydropyridin-3-yl)-2-nitrobenzenesulfonamide

A solution of Intermediate 119 (8.8 g, 14.02 mmol) in DCM (120 mL) atroom temperature was added to zinc bromide (9.47 g, 42.05 mmol). Thereaction mixture was stirred under N₂ at room temperature overnight.LCMS showed reaction was completed to give the secondary amine. It wasfiltered and washed DCM. The DCM layer was washed with aq NaHCO₃, brine,dried over MgSO₄, filtered and concentrated to give a residue, which wasadsorbed onto silica gel and purified by silica gel column (40 g, 20-80%hex/EA) to afford the title compound (5.00 g, 67.6%) as an oil.

MS: 528 ES+ (C₂₃H₃₇N₃O₇SSi)

Intermediate 121:(S)—O-allyl-N-(6-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine

To a stirred suspension of Intermediate 120 (5 g, 9.47 mmol) and K₂CO₃(7.86 g, 56.85 mmol) in acetonitrile (100 mL), PhSH (3.90 mL, 37.90mmol) was added. The reaction was stirred at rt overnight. It wasfiltered and concentrated, then diluted with DCM and filtered throughdisposal filter. The filtrate was concentrated and purified by silicagel column (0-100% Hex/EA, 80 g ISCO) to afford the title compound (2.04g, 63%) as an oil.

MS: 343 ES+ (C₁₇H₃₄N₂O₃Si)

Intermediate 122:(S)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-4-(methoxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a stirred solution of Intermediate 121 (2.04 g, 5.96 mmol) inacetonitrile (500 mL), N-ethyl-N-isopropylpropan-2-amine (4.15 mL, 23.82mmol) was added triphosgene (0.721 g, 2.38 mmol) as a solution in 20 mLCH₃CN via syringe pump (0.1 ml/min) at 0° C. The reaction was stirredand allowed to warm from 0° C. to rt overnight. LCMS showed completeconversion to product. The solution was concentrated to give a residue,which was then taken up in 150 mL EtOAc and washed with 5% citric acid,NaHCO₃, brine, dried over MgSO₄, filtered and concentrated. The residuewas purified by silica gel column (40 g, 0-60% Hex/EA) to afford thetitle compound (1.700 g, 77%) as an oil.

MS: 369 ES+ (C₁₈H₃₂N₂O₄Si)

Intermediate 123:(S)-6-(allyloxy)-2-(hydroxymethyl)-4-(methoxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a stirred solution of Intermediate 122 (1.7 g, 4.61 mmol) in THF (40mL), TBAF (6.92 mL, 6.92 mmol) was added at 0° C. The reaction mixturewas stirred at 0° C. for 1 hr. LCMS indicated the disappearance ofstarting material and formation of the desired product. The mixture wasconcentrated and purified by silica gel column (40 g, 50-100% Hex/EA) togive the title compound (1.050 g, 90%) as a white solid.

MS: 255 ES+ (C₁₂H₁₈N₂O₄)

Intermediate 124:(S)-6-(allyloxy)-2,4-bis(methoxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a stirred solution of Intermediate 123 (0.3 g, 1.18 mmol) in ACN (12mL) was added iodomethane (0.734 mL, 11.80 mmol) and silver oxide (1.094g, 4.72 mmol). The reaction was stirred at rt with aluminum foil wrappedaround overnight. LCMS showed product. The mixture was filtered andconcentrated. The residue was then purified by silica gel column (40 g,0-100% Hex/EA) to afford the title compound (0.100 g, 31.6%) as an oil.

MS: 269 ES+ (C₁₃H₂₀N₂O₄)

Example 14(2S,5R)-2-(1-(tert-butoxycarbonyl)piperidin-4-ylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate Sodium Salt

In a beaker, the DOWEX resin 50WX8 (15 g) was conditioned by stirring itfor 20 min in 2N sodium hydroxide (39.3 mL, 78.69 mmol), loaded onto acartridge, washed with water until pH equaled 7, and then was rinsedwith acetone/water (1/1) followed by water again. The phosphonium salt(Intermediate 132, 140 mg) was loaded using water and a minimum amountof acetone and eluted with water. The fractions were directly frozen andlyophilized to give the desired sodium salt as a white solid (39 mg).

MS: 445 (M-H) (C₁₇H₂₆N₄O₈S.[Na⁺])

¹H NMR (300 MHz, DMSO-d) δ: 1.31-1.40 (m, 2H) 1.40-1.45 (s, 9H) 1.70(m., 2H) 2.79-2.86 (m, 2H) 3.21 (d, 1H) 3.85 (dd, 2H) 3.75 (m, 1H)3.80-3.93 (m, 2H) 4.01-4.12 (m, 1H) 4.25 (s, 1H) 5.82 (d, 1H) 6.30-6.36(m, 1H)

Intermediate 125: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of cerious chloride (4.36 g, 11.71 mmol) and(S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 50, 4.00 g, 11.71 mmol) in 50 mL MeOH at 0° C., NaBH₄(0.443 g, 11.71 mmol) was added as a solid. The mixture was stirred atambient temp for 15 minutes. The mixture was concentrated and dilutedwith 10% citric acid (aq), H₂O and ethyl acetate. The organic layer wasseparated and washed with brine, dried over MgSO₄, filtered andconcentrated to give a residue which was purified by silica gel column(25 g, 20-40% Hex/EA) to give the desired product (2.6 g) as a colorlessoil.

MS: 344 ES+ (C₁₇H₃₃NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.10-0.07 (m, 6H) 0.65-0.88 (m, 9H) 1.25(br. s., 6H) 3.60 (d, J=5.67 Hz, 3H) 5.08 (d, J=5.10 Hz, 1H) 5.67 (br.s., 1H) 5.72-5.86 (m, 1H)

Intermediate 126: (2S,5R)-tert-butyl5-(N-(allyloxy)-2,4-dinitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 125, 17.50 g, 50.94 mmol), phosphorus triphenyl (14.70 g,56.04 mmol), N-(allyloxy)-2,4-dinitrobenzenesulfonamide (16.22 g, 53.49mmol) in toluene (380 mL) was added diisopropyl azodicarboxylate (11.03ml, 56.04 mmol) at room temperature. The reaction mixture was stirred atrt overnight. The mixture was concentrated and the residue was purifiedby silica gel column (220 g, 0-40% Hex/EA) to give the desired product(30.8 g) as a yellow gum.

MS: 429 ES+ (C₂₆H₄₀N₄O₁₀SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.10-0.07 (m, 6H) 0.65-0.88 (m, 9H) 1.25(br. s., 6H) 1.32 (s, 4H) 5.14 (d, J=10.01 Hz, 2H) 8.01 (dd, J=9.25,1.89 Hz, 4H) 8.10 (s, 1H)

Intermediate 127:N-(allyloxy)-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)-2,4-dinitrobenzenesulfonamide

To a stirred solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2,4-dinitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 126, 19.5 g, 31.01 mmol) in DCM (250 mL) under nitrogen atrt, zinc bromide (20.95 g, 93.04 mmol) was added. The reaction mixturewas stirred at rt for 6 hs, (lcms indicated the reaction was a cleanone) and diluted with 50 mL DCM and washed with sat. NaHCO₃, brine,dried over MgSO₄, filtered and concentrated to give the desired productas an orange oil (17.5 g).

MS: 529 ES+ (C₂₁H₃₂N₄O₈SSi)

¹H NMR (300 MHz, CHLOROFORM-d) δ: −0.17-0.11 (m, 6H) 0.65-0.87 (m, 9H)3.1 (m, 2H), 3.75 (m, 3H) 4.6 (m 3H) 5.26 (d, J=8.69 Hz, 2H) 5.86 (m,J=8.69 Hz, 3H) 8.17-8.57 (m, 3H)

Intermediate 128:O-allyl-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine

To a stirred suspension ofN-(allyloxy)-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)-2,4-dinitrobenzenesulfonamide(Intermediate 127, 17.50 g, 33.10 mmol) and potassium carbonate (13.73g, 99.31 mmol) in acetonitrile (700 mL), benzenethiol (5.10 ml, 49.65mmol) was added. The reaction mixture was stirred at rt for 2.5 hours.LCMS showed the completion of the reaction. The reaction wasconcentrated, the diluted with EtOAc and filtered through a disposalfilter. The filtrate was concentrated and the crude product was purifiedon silica gel column (0-4% MeOH in DCM) to get the desired product as ayellow oil (5.6 g).

MS: 299 ES+ (C₁₅H₃₀N₂O₂Si)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.08-0.09 (m, 6H) 0.75-0.92 (m, 9H)2.53-2.62 (m, 1H) 3.03 (dd, J=11.71, 4.72 Hz, 1H) 3.15-3.25 (m, 1H) 3.29(s, 1H) 3.37 (td, J=4.58, 2.36 Hz, 1H) 3.41-3.49 (m, 2H) 4.08 (dt,J=5.57, 1.37 Hz, 2H) 5.04-5.27 (m, 2H) 5.61-5.80 (m, 2H) 5.80-6.00 (m,1H) 6.40 (d, J=7.37 Hz, 1H)

Intermediate 129:(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a stirred solution ofO-allyl-N-((3R,6S)-6-((tert-butyldimethylsilyloxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine(Intermediate 128, 2.0 g, 6.70 mmol) in acetonitrile (560 mL),N-ethyl-N-isopropylpropan-2-amine (4.67 ml, 26.80 mmol) was added.Bis(trichloromethyl) carbonate (0.795 g, 2.68 mmol) dissolved in 40 mLCH₃CN was added via a syringe pump (at a rate 0.1 ml/min) at 0° C. Thereaction mixture was stirred at 0° C. for 1 hr. It was then allowed tostir at room temperature overnight. The mixture was then concentratedand diluted with EtOAc (50 mL). The organic layer was washed with water,brine, dried over MgSO₄, filtered and concentrated. The residue waspurified by silica gel column (12 g, 0-25% Hex/EA) to give the desiredproduct as a colorless oil (1.64 g).

MS: 325 ES+ (C₁₆H₂₈N₂O₃Si)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.05-0.07 (m, 6H) 0.67-0.87 (m, 9H)2.92-3.08 (m, 1H) 3.32 (d, J=11.14 Hz, 1H) 3.48-3.62 (m, 1H) 3.68-3.82(m, 2H) 3.87 (dd, J=5.00, 2.74 Hz, 1H) 4.29 (dt, J=6.00, 1.25 Hz, 2H)5.09-5.39 (m, 2H) 5.50-6.02 (m, 2H) 6.11-6.39 (m, 1H)

Intermediate 130:(2S,5R)-6-(allyloxy)-2-(hydroxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a stirred solution of(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 129, 1.64 g, 5.05 mmol) in THF (40 mL), tetrabutylammoniumfluoride (6.06 ml, 6.06 mmol) was added at 0° C. The reaction mixturewas stirred at 0° C. for 1 hr. The mixture was concentrated and purifiedby silica gel column (40 g, 50-100% Hex/EA) to give the desired product(1.03 g) as an yellow oil.

MS: 211 ES+ (C₁₀H₁₄N₂O₃)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 3.31-3.37 (m, 2H) 3.69-3.81 (m, 2H)3.88 (dd, J=4.34, 1.51 Hz, 1H) 3.97-4.11 (m, 1H) 4.46 (ddt, J=7.86,6.63, 1.06, 1.06 Hz, 2H) 5.24-5.45 (m, 2H) 5.56-5.72 (m, 1H) 5.92-6.18(m, 1H) 6.30-6.49 (m, 1H)

Intermediate 131: tert-butyl4-((2S,5R)-6-(allyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-enecarboxamido)piperidine-1-carboxylate

To a solution of periodic acid (6 g, 31.26 mmol) in wet acetonitrile (60mL) (0.75% water by volume) at room temperature was added chromium(VI)oxide (10 mg, 0.10 mmol). The mixture was stirred until completedissolution was achieved. This orthoperiodic acid solution was used inthe next step.

To a stirred solution of(2S,5R)-6-(allyloxy)-2-(hydroxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 130, 216 mg, 1.03 mmol) in wet CH₃CN (10 mL) (0.75% waterby volume) at rt, 9 mL previously formed orthoperiodic acid solution wasadded dropwise. The mixture was then stirred at rt for 50 minutes, when50 mL of EtOAc was added. The organic solution was washed with smallamount of water and brine then dried over MgSO₄, filtered, concentratedto give a light yellow oil as the crude acid (200 mg). To a solution ofthe crude acid in DMF (5 mL) at 0° C. was added, tert-butyl4-aminopiperidine-1-carboxylate (214 mg, 1.07 mmol),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (509 mg, 1.34 mmol) and TEA (497 μl, 3.57 mmol). Thereaction was stirred at 0° C. for 35 min, then the mixture was dilutedwith ethyl acetate and washed with brine and water. The combined aqueouswashes were extracted once with ethyl acetate. The combined organicextracts were dried over magnesium sulfate, filtered and concentrated.Silica gel chromatography (12 g column, 50%-100% EtOAc) afforded thedesired product as a colorless oil (110 mg).

MS: 407 ES+ (C₂₀H₃₀N₄O₅)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.31-1.43 (m, 2H) 1.42-1.51 (m, 9H)1.90 (br. s., 2H) 2.79-2.96 (m, 2H) 3.01 (d, J=10.39 Hz, 1H) 3.38 (dd,J=10.48, 1.61 Hz, 1H) 3.85 (dd, J=5.00, 3.12 Hz, 1H) 3.89-3.98 (m, 1H)4.01-4.12 (m, 2H) 4.35-4.52 (m, 3H) 5.22-5.45 (m, 2H) 5.93-6.08 (m, 1H)6.15 (dd, J=9.35, 3.30 Hz, 1H) 6.32-6.50 (m, 1H) 6.84 (d, J=7.55 Hz, 1H)

Intermediate 132: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(1-(tert-butoxycarbonyl)piperidin-4-ylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

To a solution of tert-butyl4-((2S,5R)-6-(allyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-enecarboxamido)piperidine-1-carboxylate(Intermediate 131, 110 mg, 0.52 mmol) in DCM (5 mL) was added AcOH (59.2μl, 1.03 mmol) (dried over Na₂SO₄) andtetrakis(triphenyl-phosphine)palladium(0) (597 mg, 0.52 mmol) undernitrogen at room temperature. The yellow solution was stirred at roomtemperature for 1 hr. To the mixture was added pyridine (10 mL),followed by pyridine/sulfur trioxide complex (493 mg, 3.10 mmol). Thesuspension was stirred at room temperature overnight under nitrogen. Thereaction mixture was concentrated to dryness (≦30° C.) and then purifiedon silica gel column (acetone in hexane (5 to 100%)) to yield anoff-white solid (140 mg).

Example 15(2S,5R)-4-(dimethylcarbamoyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate Sodium Salt

Dowex(R) 50WV8, 100-200 mesh, ion exchange resin (0.63 g, 9.41 μmol) wasconditioned by stirring for 3 hours in 2N NaOH (1.6 mL, 3.20 mmol). Theresin was then loaded into a cartridge and washed with water until thepH was 7. It was then washed with (1/1) acetone/water, followed bywater. (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-4-(dimethylcarbamoyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 144, 6 mg, 9.41 μmol) was taken up in water. Theyellow solution was loaded on the resin and washed through with water.The product was collected. Dried on lyophilizer to obtain the desiredproduct (3.3 mg, 98%) as an off-white solid.

MS: 334 ES− (C₁₁H₁₆N₃O₇SNa)

¹H NMR (300 MHz, D₂O) δ: 2.99 (s, 7H) 3.13 (s, 7H) 3.44 (s, 6H) 3.56 (d,J=1.70 Hz, 4H) 3.65-3.79 (m, 5H) 4.24 (ddd, J=8.34, 4.47, 3.01 Hz, 2H)4.47 (s, 2H) 5.97 (d, J=1.88 Hz, 2H)

Intermediate 133: (2S,5R)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

A dry, two-necked, round-bottomed flask equipped with a reflux condenserfitted with a nitrogen inlet at its top, a rubber septum, and a magneticstirring bar was charged with toluene (7.5 mL) and flushed briefly withnitrogen after which trimethylaluminum in hexanes (3.5 mL, 7.0 mmol) wasinjected through the septum into the flask. The solution was stirred andcooled in an ice-salt bath at −10° to −15°, and dimethylamine in THF(3.36 mL, 6.72 mmol) was added slowly with a syringe. Twenty minutesafter the addition was completed, the cooling bath was removed, and thecontents of the flask were allowed to stir and warm slowly to roomtemperature over a 45-minute period. A solution of (2S,5R)-1-tert-butyl4-methyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1,4(2H)-dicarboxylate(Intermediate 102, 2.25 g, 5.60 mmol) in toluene (3 mL) was prepared andinjected into two-necked flask through the septum under nitrogen,immediate bubbling was observed. The resulting solution was stirredovernight at rt. LCMS showed a tiny amount of starting material left.Another batch of Me₂AlNMe₂ was prepared using 0.6 mL amine and 0.6 mLAlMe₃ in 1 mL toluene and transferred into reaction mix. The reactionwas stirred for 3 more hours. The reaction was then hydrolyzed by slow,cautious addition of 7.82 mL (7.82 mmole) of 1M HCl. The mixture wasthen stirred for 30 minutes to ensure complete hydrolysis. The upperorganic layer was separated, and the aqueous layer was extracted withthree 50 mL portions of EtOAc. The organic extracts were combined,washed with brine, dried with anhydrous Na₂SO₄ and evaporated underreduced pressure to obtain the desired product (2.194 g, 94%) as a clearbrown oil.

MS: 415 ES+ (C₂₀H₃₈N₂O₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (s, 6H) 0.83-0.86 (m, 9H) 1.40-1.44(m, 9H) 2.91 (br. s., 7H) 3.66-3.73 (m, 2H) 4.04 (m, J=7.00 Hz, 1H)4.15-4.22 (m, 1H) 4.33-4.51 (m, 1H) 4.98 (d, J=6.22 Hz, 1H) 5.88 (d,J=3.58 Hz, 1H)

Intermediate 134: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 133, 2.194 g, 5.29 mmol) in DCM (65 mL) was cooled in anice-bath then Dess-Martin periodinane in DCM (26.5 mL, 7.94 mmol) wasadded and the reaction was stirred at rt overnight. The organic solutionwas washed with sat. Na₂S₂O₃, sat. NaHCO₃, brine, dried over MgSO₄,filtered and evaporated. The crude product was purified via flashchromatography (10-75% EA/EtOAc) to obtain the desired product (1.050 g,48.1%).

MS: 413 ES+ (C₂₀H₃₆N₂O₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (s, 6H) 0.83 (s, 9H) 1.44 (s, 9H) 2.81(s, 3H) 2.89 (s, 3H) 3.79-4.02 (m, 3H) 4.30-4.48 (m, 1H) 4.72-4.87 (m,1H) 7.10-7.22 (m, 1H)

Intermediate 135: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of cerium(III) chloride heptahydrate (0.948 g,2.54 mmol) and (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 134, 1.05 g, 2.54 mmol) in 1 mL MeOH at 0° C., sodiumtetrahydroborate (0.096 g, 2.54 mmol) was added as a solid. The mixturewas stirred at ambient temp for 15 minutes. The mixture was concentratedand diluted with NH₄Cl (aq), H₂O and ether. The ether layer wasseparated and washed with brine, dried over Na₂SO₄, filtered andconcentrated to give the desired product (1.034 g, 98%) as a beigesolid.

MS: 415 ES+ (C₂₀H₃₈N₂O₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H) 0.76-0.92 (m, 9H) 1.30-1.48(m, 9H) 2.71-3.05 (m, 7H) 3.74 (d, J=4.71 Hz, 2H) 4.18 (br. s., 3H) 5.34(d, J=5.46 Hz, 1H) 5.66 (br. s., 1H)

Intermediate 136:(3S,6S)-1-(tert-butoxycarbonyl)-6-((tert-butyldimethylsilyloxy)methyl)-3-hydroxy-1,2,3,6-tetrahydropyridine-4-carboxylicacid

(2S,5S)-1-tert-butyl 4-methyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1,4(2H)-dicarboxylate(Intermediate 101, 4.22 g, 10.51 mmol) was dissolved in THF (64.0 mL)and water (32 mL) and then LiOH (0.277 g, 11.56 mmol) was added andstirred at rt overnight. The reaction mixture was acidified with 1N HCl,extracted with EtOAc, washed with brine, dried over Na₂SO₄, filtered andconcentrated to obtain the desired product (4.07 g, 100%) as atransparent oil.

MS: 386 ES− (C₁₈H₃₃NO₆Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.04 (s, 6H) 0.84-0.90 (m, 9H) 1.40 (s, 9H)2.73-2.88 (m, 1H) 3.75 (d, J=5.46 Hz, 2H) 4.25 (t, J=7.16 Hz, 1H) 4.41(br. s., 1H) 5.15 (br. s., 1H) 6.65 (d, J=3.96 Hz, 1H) 12.46 (br. s.,1H)

Intermediate 137: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of(3S,6S)-1-(tert-butoxycarbonyl)-6-((tert-butyldimethylsilyloxy)methyl)-3-hydroxy-1,2,3,6-tetrahydropyridine-4-carboxylicacid (Intermediate 136, 4.07 g, 10.50 mmol) in DMF (20 mL) at roomtemperature was added 2M dimethylamine in THF (5.25 mL, 10.50 mmol),HATU (5.99 g, 15.75 mmol) and then DIEA (5.50 mL, 31.51 mmol). Thereaction mixture was stirred at room temperature. The reaction ran untilthe starting material was consumed, then it was quenched with water andextracted with EtOAc. The organic solutions was then washed with brine,and purified via flash chromatography (35-100% EA/Hex) to isolate thedesired product (3.05 g, 70.0%) as a pink glassy solid.

MS: 415 ES+ (C₂₀H₃₈N₂O₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H) 0.76-0.92 (m, 9H) 1.30-1.48(m, 9H) 2.71-3.05 (m, 7H) 3.74 (d, J=4.71 Hz, 2H) 4.18 (br. s., 3H) 5.34(d, J=5.46 Hz, 1H) 5.66 (br. s., 1H)

Intermediate 138: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5,6-dihydropyridine-1(2H)-carboxylate

A stirred suspension of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(dimethylcarbamoyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 137, 3.09 g, 7.45 mmol),N-(allyloxy)-2-nitrobenzenesulfonamide (2.348 g, 9.09 mmol) andtriphenylphosphine (2.150 g, 8.20 mmol) in toluene (65 mL) was cooled ina salt-ice-bath and then (E)-diisopropyl diazene-1,2-dicarboxylate(1.588 mL, 8.20 mmol) was added dropwise. The reaction was let warm upto rt and was stirred for an additional 2 h. The solvent was removed andthe crude product was loaded onto silica gel, purified via flashchromatography (25-75% EA/Hex) to obtain the desired product (4.0 g,82%) as an off-white solid.

MS: 655 ES+ (C₂₉H₄₆N₄O₉SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.01 (s, 6H) 0.82 (s, 9H) 1.37 (d, J=12.24Hz, 9H) 2.56-3.06 (m, 6H) 3.06-3.25 (m, 1H) 3.60-3.81 (m, 2H) 4.07-4.41(m, 3H) 4.41-4.68 (m, 1H) 4.73 (br. s., 1H) 5.14-5.33 (m, 2H) 5.70-5.92(m, 1H) 6.18-6.31 (m, 1H) 7.83-8.09 (m, 4H)

Intermediate 139: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(dimethylcarbamoyl)-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethyl-silyloxy)methyl)-4-(dimethylcarbamoyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 138, 4 g, 6.11 mmol) in THF (20 mL) was charged withnitrogen and cooled in an ice-bath. To this solution was added 1M TBAFin THF (7.33 mL, 7.33 mmol) dropwise and stirred for 1 h. The solventwas then removed and the crude product loaded onto silica gel andpurified via flash chromatography (0-20% MeOH-DCM) to obtain the desiredproduct (2.98 g, 90%) as a beige solid.

MS: 541 ES+ (C₂₃H₃₂N₄O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.38 (d, J=6.03 Hz, 9H) 2.85 (br. s., 6H)3.04-3.27 (m, 1H) 3.51 (br. s., 2H) 4.27 (br. s., 3H) 4.39-4.64 (m, 1H)4.75 (br. s., 1H) 4.87-4.99 (m, 1H) 5.24 (t, J=3.96 Hz, 2H) 5.73-5.91(m, 1H) 6.18-6.35 (m, 1H) 8.00 (br. s., 4H)

Intermediate 140: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(dimethylcarbamoyl)-2-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(dimethylcarbamoyl)-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 139, 1.64 g, 3.03 mmol) and silver oxide (2.81 g, 12.13mmol) were dissolved in acetonitrile (40 mL). To this solution was addediodomethane (1.897 mL, 30.34 mmol) under N₂ and stirred over for 5 daysprotected from the light. The reaction mixture was then filtered throughCelite, rinsed with EtOAc, and the crude product was loaded onto silicagel. The product was purified by flash chromatography (0-20% MeOH/DCM)to yield the desired product (1.643 g, 98%) as an off-white solid.

MS: 555 ES+ (C₂₄H₃₄N₄O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.38 (d, J=7.35 Hz, 9H) 2.57-3.20 (m, 7H)3.22 (s, 3H) 3.44 (d, J=3.58 Hz, 2H) 4.08-4.35 (m, 3H) 4.57-4.82 (m, 2H)5.17-5.31 (m, 2H) 5.73-5.90 (m, 1H) 6.16-6.32 (m, 1H) 7.82-7.92 (m, 1H)7.99 (d, J=6.22 Hz, 3H)

Intermediate 141:(3R,6S)-3-(N-(allyloxy)-2-nitrophenylsulfonamido)-6-(methoxymethyl)-N,N-dimethyl-1,2,3,6-tetrahydropyridine-4-carboxamide

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(dimethylcarbamoyl)-2-(methoxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 140, 1.642 g, 2.96 mmol) in DCM (8 mL) was addedtrifluoroacetic acid (4.33 mL, 56.25 mmol) and stirred at rt for 30minutes. Upon addition, the color of the solution immediately turnedpink. Then the solvents were evaporated and the crude productredissolved in DCM and washed with 0.5N NaOH, brine, filtered andconcentrated to obtain the desired product (1.346 g, 100%) as a beigefoam.

MS: 455 ES+ (C₁₉H₂₆N₄O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.62-3.03 (m, 9H) 3.20-3.28 (m, 5H) 3.55(br. s., 1H) 4.31-4.44 (m, 1H) 4.48 (d, J=6.97 Hz, 1H) 4.65 (br. s., 1H)5.19-5.33 (m, 2H) 5.81-5.97 (m, 1H) 6.12 (s, 1H) 7.86-7.92 (m, 1H)7.95-8.09 (m, 2H)

Intermediate 142:(3R,6S)-3-(allyloxyamino)-6-(methoxymethyl)-N,N-dimethyl-1,2,3,6-tetrahydropyridine-4-carboxamide

To a solution of(3R,6S)-3-(N-(allyloxy)-2-nitrophenylsulfonamido)-6-(methoxymethyl)-N,N-dimethyl-1,2,3,6-tetrahydropyridine-4-carboxamide(Intermediate 141, 1.346 g, 2.96 mmol) and potassium carbonate (2.456 g,17.77 mmol) in acetonitrile (50 mL) was added benzenethiol (1.825 mL,17.77 mmol) and stirred at rt overnight. The solvent was evaporated thenthe crude mixture was redissolved in DCM and a little bit MeOH,filtered, loaded onto silica gel. The crude product was purified viaflash chromatography (0-20% MeOH/DCM) and dried under high-vacuum toobtain the desired product (0.725 g, 91%) as a pale yellow oil.

MS: 270 ES+ (C₁₃H₂₃N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.78-3.00 (m, 9H) 3.20-3.28 (m, 5H) 3.46(br. s., 1H) 3.58 (d, J=8.10 Hz, 1H) 4.02 (dt, J=5.51, 1.39 Hz, 2H)5.08-5.23 (m, 2H) 5.79-5.92 (m, 2H) 6.36-6.41 (m, 1H)

Intermediate 143:(2S,5R)-6-(allyloxy)-2-(methoxymethyl)-N,N-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide

A solution of(3R,6S)-3-(allyloxyamino)-6-(methoxymethyl)-N,N-dimethyl-1,2,3,6-tetrahydropyridine-4-carboxamide(Intermediate 142, 725 mg, 2.69 mmol) and Hunig's Base (1.880 mL, 10.77mmol) in acetonitrile (350 mL) was cooled to below 0° C. in an ice-saltbath and then a solution of triphosgene (320 mg, 1.08 mmol) in ACN (22mL) was added at a rate of 0.1 mL/min. The reaction was stirredovernight at rt. Upon completion of the reaction, the solvents wereremoved and the crude mixture was redissolved in EtOAc, washed with sat.NaHCO₃ and brine, dried over MgSO₄, filtered and concentrated to obtainthe desired product (658 mg, 83%) as a pale yellow oil.

MS: 296 ES+ (C₁₄H₂₁N₃O₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.92 (br. s., 6H) 3.16-3.22 (m, 1H) 3.28(s, 3H) 3.32-3.38 (m, 1H) 3.51-3.65 (m, 2H) 3.92 (ddd, J=6.40, 4.99,2.92 Hz, 1H) 4.16 (d, J=2.64 Hz, 1H) 4.34 (dd, J=5.84, 1.32 Hz, 2H)5.20-5.35 (m, 2H) 5.80 (dd, J=2.92, 1.04 Hz, 1H) 5.81-5.95 (m, 1H)

Intermediate 144: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-4-(dimethylcarbamoyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

To a solution of(2S,5R)-6-(allyloxy)-2-(methoxymethyl)-N,N-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide(Intermediate 143, 115 mg, 0.39 mmol) and AcOH (0.045 mL, 0.78 mmol)(dried over sodium sulfate) in CH₂Cl₂ (3 mL) at room temperature wasadded Pd(Ph₃P)₄ (450 mg, 0.39 mmol) under a nitrogen atmosphere. Thesolution was stirred at rt for 1 h. To this reaction was added pyridine(3.00 mL) and sulfur trioxide pyridine complex (496 mg, 3.12 mmol) andcontinued stirring under N₂ at rt overnight. The desired mass wasobserved by LC/MS in the reaction mixture. The suspension was evaporatedto dryness and then resuspended in DCM. The solids were filtered offthrough a 0.2μ nalgene filter. The filtrate was concentrated to afford ayellow oil. The 80 mg crude was purified using prep HPLC usingWater/Formic Acid. The product was then lyophilized to obtain 6 mgslightly impure product which was kept under inert atmosphere until nextstep.

Example 16(2S,5R)-2-(hydroxymethyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

In a 150 mL of beaker, the DOWEX resin 50WX8 (50 g) was conditioned bystirring it for 20 min in 2N sodium hydroxide (40 mL), loaded onto acartridge, washed with water until pH was 7, then washed withacetone/water (1/1) and water again.(2S,5R)-2-((tert-butyldimethylsilyloxy)-methyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate-(E)-triphenyl(prop-1-enyl)phosphonium (Intermediate 145, 190 mg)was loaded using water and a minimum amount of acetone and eluted withwater. The fractions were directly frozen and lyophilized to give thecompound as a white solid (130 mg), which was purified by a reversedphase chromatography (MeCN in water: 0-4%) to give a white solid as thedesired product (31 mg).

MS: 263 ES− (C8H12N2O6S.[Na+])

¹H NMR (300 MHz, DMSO-d₆) δ: 1.76 (t, J=1.70 Hz, 3H) 3.02-3.14 (m, 1H)3.15-3.25 (m, 1H) 3.41-3.52 (m, 1H) 3.52-3.62 (m, 2H) 3.92 (d, J=2.83Hz, 1H) 4.72-4.89 (m, 1H) 5.27 (s, 1H)

Intermediate 145:(2S,5R)-2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate-(E)-triphenyl(prop-1-enyl)phosphonium

To a solution of(2S,5R)-6-(allyloxy)-2-((tert-butyldimethylsilyloxy)methyl)-4-methyl-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 13, 0.40 g, 1.18 mmol) in DCM (20 mL) was added driedacetic acid (0.135 mL, 2.36 mmol) andtetrakis(triphenylphosphine)palladium(0) (1.365 g, 1.18 mmol) undernitrogen at room temperature. The yellow solution was stirred at roomtemperature for 4 hours, LC/MS indicated the starting material was gone.Then to the mixture was added pyridine (20 ml), followed bypyridine-sulfurtrioxide (1.128 g, 7.09 mmol). The suspension was stirredat room temperature overnight under nitrogen. The mixture wasconcentrated to dryness (<=30° C.), redissolved in DCM and filtered. Thecrude product was purified on silica gel column (Acetone in DCM: 10-40%)to give a white solid (190 mg) as the desired product.

MS: 377 ES− (C₁₄H₂₅N₂O₆SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: ppm −0.11-0.09 (m, 6H) 0.73-0.88 (m, 9H)1.21-1.22 (m, OH) 1.71 (t, J=1.89 Hz, 3H) 2.06-2.19 (m, 2H) 3.03 (dd,J=11.05, 2.93 Hz, 1H) 3.42-3.57 (m, 1H) 3.60-3.79 (m, 2H) 3.88 (d,J=3.02 Hz, 1H) 5.20 (dt, J=2.74, 1.46 Hz, 1H) 6.44-6.73 (m, 1H)7.08-7.36 (m, 1H) 7.51-7.79 (m, 10H) 7.80-7.98 (m, 3H)

Example 17(2S,5R)-3-methyl-7-oxo-2-(piperidin-1-ium-4-ylcarbamoyl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

To a solution of (E)-triphenyl(prop-1-en-1-yl)phosphonium(2S,5R)-2-((1-(tert-butoxycarbonyl)piperidin-4-yl)carbamoyl)-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 152, 0.2 g, 0.26 mmol) in DCM (3 mL) at 0° C. wasadded TFA (0.5 mL). The reaction mixture was stirred for 30 minutes andconcentrated to afford a yellow oil. The residue was redissolved in DCMand extracted with water. The aqueous was lyophilized. Purification wasdone on reverse phase HPLC (0%-20% acetonitrile in water, Synergi PolarRP 21.2 mm×100 mm, 4 μm coupled with YMC C30, 20 mm×150 mm, 5 μm) toafford the title compound as a white solid (18.7 mg, 20%).

Optical Rotation: (0.16 g/dL, DMSO)=−291

MS: 359 ES− (C₁₃H₂₀N₄O₆S)

¹H NMR (300 MHz, D₂O) δ: 1.74 (m, 3H); 1.82 (m, 2H); 2.19 (m, 2H); 3.15(m, 2H); 3.47 (m, 3H); 3.61 (m, 1H); 4.05 (m, 1H); 4.29 (m, 1H); 4.39(m, 1H); 6.30 (m, 1H).

Intermediate 146:N-[(3R,6S)-6-[[(tert-butyldimethylsilyl)oxy]methyl]-5-methyl-1,2,3,6-tetrahydropyridin-3-yl]-2-nitro-N-(prop-2-en-1-yloxy)benzene-1-sulfonamide

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution of tert-butyl(3R,6S)-6-[[(tert-butyldimethylsilyl)oxy]methyl]-5-methyl-3-[N-(prop-2-en-1-yloxy)(2-nitrobenzene)sulfonamido]-1,2,3,6-tetrahydropyridine-1-carboxylate(Intermediate 80, 13.6 g, 22.75 mmol, 1.00 equiv) in dichloromethane(100 mL). This was followed by the addition of ZnBr₂ (10.2 g, 45.29mmol, 2.00 equiv) in several batches. The resulting solution was stirredovernight at room temperature. The resulting solution was diluted with500 mL of dichloromethane. The resulting mixture was washed with 2×200mL of sodium bicarbonate (aq) and 2×200 mL of NH₄Cl (aq). The mixturewas dried over anhydrous sodium sulfate and concentrated under vacuum.This resulted in 12 g (crude product) of the title compound as yellowoil.

MS: 498 ES+ (C₂₂H₃₅N₃O₆SSi)

Intermediate 147:(3R,6S)-6-[[(tert-butyldimethylsilyl)oxy]methyl]-5-methyl-N-(prop-2-en-1-yloxy)-1,2,3,6-tetrahydropyridin-3-amine

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed a solution ofN-[(3R,6S)-6-[[(tert-butyldimethylsilyl)oxy]methyl]-5-methyl-1,2,3,6-tetrahydropyridin-3-yl]-2-nitro-N-(prop-2-en-1-yloxy)benzene-1-sulfonamide(Intermediate 146, 12 g, 24.11 mmol, 1.00 equiv) inN,N-dimethylformamide (100 mL), 2-sulfanylacetic acid (4.4 g, 47.77mmol, 2.00 equiv). This was followed by the addition of LiOH (5.8 g,242.17 mmol, 10.00 equiv), in portions. The resulting solution wasstirred for 2 h at room temperature. The resulting solution was dilutedwith 500 mL of water. The resulting solution was extracted with 5×200 mLof ethyl acetate and the organic layers combined. The resulting mixturewas washed with 3×200 mL of brine and 2×200 mL of sodium bicarbonate(aq.). The mixture was dried over anhydrous sodium sulfate andconcentrated under vacuum. This resulted in 8.4 g (crude product) of thetitle compound as yellow oil.

MS: 313 ES+ (C₁₆H₃₂N₂O₂Si)

Intermediate 148:(2S,5R)-2-[[(tert-butyldimethylsilyl)oxy]methyl]-3-methyl-6-(prop-2-en-1-yloxy)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

Into a 2000-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen, was placed a solution of(3R,6S)-6-[[(tert-butyldimethylsilyl)oxy]methyl]-5-methyl-N-(prop-2-en-1-yloxy)-1,2,3,6-tetrahydropyridin-3-amine(Intermediate 147, 8.4 g, 26.88 mmol, 1.00 equiv) in acetonitrile (1600mL), N,N-Diisopropylethylamine (14.2 g, 109.87 mmol, 4.00 equiv). Thiswas followed by the addition of a solution of ditrichloromethylcarbonate (2.9 g, 9.77 mmol, 0.40 equiv) in acetonitrile (100 mL)dropwise with stirring at −15° C. in 3 hr. The resulting solution wasstirred overnight at room temperature. The resulting mixture wasconcentrated under vacuum. The resulting solution was diluted with 500mL of ethyl acetate. The resulting mixture was washed with 2×400 mL ofNH₄Cl (aq.) and 2×400 mL of brine. The resulting mixture wasconcentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (1:10). This resulted in 3.9 g(43%) of the title compound as yellow oil.

MS: 339 ES+ (C₁₇H₃₀N₂O₃Si)

Intermediate 149:(2S,5R)-2-(hydroxymethyl)-3-methyl-6-(prop-2-en-1-yloxy)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

Into a 100-mL round-bottom flask, was placed tetrahydrofuran (30 mL),(2S,5R)-2-[[(tert-butyldimethylsilyl)oxy]methyl]-3-methyl-6-(prop-2-en-1-yloxy)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 148, 3.2 g, 9.45 mmol, 1.00 equiv) and it was cooled to 0°C., then TBAF (14.2 mL 1N in THF, 1.50 equiv) was added dropwise. Theresulting solution was stirred for 1 h at 0° C. in a water/ice bath. Theresulting mixture was concentrated under vacuum. The residue was appliedonto a silica gel column with ethyl acetate/petroleum ether (1:5-1:2).This resulted in 1.6 g (75%) of the title compound as a light yellowsolid.

MS: 225 ES+ (C₁₁H₁₆N₂O₃)

¹H NMR (300 MHz, CDCl₃) δ 1.63 (3H, d), 3.20 (2H, d), 3.62-3.84 (2H, m),3.85-3.90 (2H, m), 4.35-4.48 (2H, m), 5.28-5.39 (2H, m), 5.95-6.08 (2H,m).

Intermediate 150:2S,5R)-3-methyl-7-oxo-6-(prop-2-en-1-yloxy)-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid

To a solution of H₅IO₆ (12.93 g, 56.71 mmol) in wet CH₃CN (150 mL, 0.75%H₂O v/v) at r.t. was added CrO₃ (128 mg, 1.28 mmol). The mixture wasstirred until complete dissolved was achieved. Into a 100-mLround-bottom flask, was placed wet acetonitrile (35 mL),(2S,5R)-2-(hydroxymethyl)-3-methyl-6-(prop-2-en-1-yloxy)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 149, 740 mg, 3.30 mmol, 1.00 equiv) and it was cooled to0° C., then the above oxidation solution (35 mL, 3.00 equiv) was addeddropwise during 30 min at 0° C. The resulting solution was stirred for 1h at 0° C. in a water/ice bath and monitor by TLC until the material wasconsumed completely. Then it was diluted with 200 mL of chloroform and50 mL of citric acid solution (25%). Separating the organic layer andthe organic layer was washed by 3×50 mL of brine, dried over anhydroussodium sulfate and concentrated under vacuum. This resulted in 0.700 gcrude of the title compound as yellow oil.

MS: 239 ES+ (C₁₁H₁₆N₂O₄)

Intermediate 151: tert-butyl4-[(2S,5R)-3-methyl-7-oxo-6-(prop-2-en-1-yloxy)-1,6-diazabicyclo[3.2.1]oct-3-ene-2-amido]piperidine-1-carboxylate

Into a 100-mL round-bottom flask, was placed N,N-dimethylformamide (20mL), crude of(2S,5R)-3-methyl-7-oxo-6-(prop-2-en-1-yloxy)-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid (Intermediate 150, 700 mg, 2.94 mmol, 1.00 equiv), tert-butyl4-aminopiperidine-1-carboxylate (1.17 g, 5.84 mmol, 1.99 equiv), HATU(1.67 g, 4.39 mmol, 1.49 equiv), DIEA (1.51 g, 11.68 mmol, 3.98 equiv)at 0° C. The resulting solution was stirred for 12 h at 25° C. Theresulting solution was diluted with 150 mL of ethyl acetate and it waswashed with 3×100 mL of brine, dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was applied onto a silica gelcolumn with ethyl acetate/petroleum ether (1:5-1:2). This resulted in0.500 g (40%) of the title compound as a white solid.

MS: =421 ES+ (C₂₁H₃₂N₄O₅)

¹H NMR (300 MHz, CDCl₃) δ 1.36 (2H, m), 1.46 (9H, s), 1.88-2.05 (5H, m),2.90 (2H, m), 3.07 (1H, m), 3.24 (1H, m), 3.79-3.92 (2H, m), 4.06 (2H,m), 4.23 (1H, s), 4.44 (2H, m), 5.29-5.39 (2H, m), 5.96-6.10 (2H, m),6.73 (1H, d).

Intermediate 152: (E)-triphenyl(prop-1-en-1-yl)phosphonium(2S,5R)-2-((1-(tert-butoxycarbonyl)piperidin-4-yl)carbamoyl)-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from tert-butyl4-((2S,5R)-6-(allyloxy)-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamido)piperidine-1-carboxylate(Intermediate 151, 0.763 g, 1.81 mmol) following the procedure describedfor Intermediate 17. Silica gel chromatography (0%-5%methanol/dichloromethane) afforded the desired product as a yellow foam(1.22 g, 88%).

MS: 459 ES−, 303 ES+ (C₁₈H₂₇N₄O₈S, C₂₁H₂OP)

Example 18(2S,5R)-2-carbamoyl-3-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from hydrogen(2S,5R)-2-carbamoyl-3-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 167, solution from ion exchange column) accordingto the procedure for Example 1. The desired product was obtained as anoff-white fluffy solid, 5.6 mg.

The route described in this example may be used to synthesize othercompounds of the invention, such as the compound described in Example10.

MS: 292 ES− (C₈H₁₀N₃O₇S)

¹H NMR (300 MHz, DEUTERIUM OXIDE-d) δ: 3.44-3.58 (m, 2H) 4.14 (s, 2H)4.37 (d, J=5.09 Hz, 1H) 4.63 (s, 1H) 6.52 (d, J=4.90 Hz, 1H).

The Intermediates for Example 18 were Prepared as Follows Intermediate153: tert-butyl benzyloxycarbamate

To a solution of O-benzylhydroxylamine hydrochloride (225 g, 1.44 mol)in dichloromethane (300 mL) was added a solution of sodium bicarbonate(261 g, 3.11 mol, 300 ml). After 1 hour the di-tert-butyl dicarbonate(375 g, 1.72 mol) was added at 0° C. The resulting solution was stirredfor 60 min at 0° C. in a water/ice bath and then the reaction wasstirred for 16 h at room temperature. The reaction was then quenched bythe addition of 300 mL of aqueous sodium bicarbonate. The aqueous phasewas extracted with 3×500 mL of dichloromethane and the organic layerswere combined and dried over anhydrous sodium sulfate and concentratedunder vacuum. Flash chromatograph on silica gel (PE: EA=10:1) afforded220 g (69%) of the title compound as a yellow oil.

¹H NMR (300 MHz, CDCl₃) δ: 1.43 (9H, s), 4.85 (2H, s), 7.19-7.21 (1H,brs), 7.30-7.40 (5H, m).

Intermediate 154: (R)-tert-butylbenzyloxy(1-hydroxybut-3-en-2-yl)carbamate

In a 3 L RBF, a solution of tert-butyl benzyloxycarbamate (Intermediate153, 86.9 g, 389.22 mmol),N,N(1S,2S)-cyclohexane-1,2-diyl)bis(2-(diphenylphosphino)-1-naphthamide)(18.5 g, 23.39 mmol), Pd₂(dba)₃-CHCl₃ (8.05 g, 7.78 mmol) and TBAB(150.5 g, 466.81 mmol) in acetonitrile (1.8 L) was bubbled with N₂ for30 minutes and the flask was closed with a rubber septum. The solutionwas then kept in a −20° C. freezer for 2 hrs. 2-vinyloxirane (30 g,428.02 mmol, 1.10 equiv) was added and the mixture was stirred in thefreezer (−20-25° C.) for 3 days. The mixture was filtered and thefiltrate was evaporated. The residue was purified by silica gel columneluted with EtOAc/hexane (0-40%) to give the title compound (106 g, 93%)as a pale yellow oil.

MS: 194 ES+ (C₁₆H₂₃NO₄)

Intermediate 155: (R)-tert-butylbenzyloxy(1-(1,3-dioxoisoindolin-2-yl)but-3-en-2-yl)carbamate

A suspension of (R)-tert-butylbenzyloxy(1-hydroxybut-3-en-2-yl)carbamate (Intermediate 154, 74 g,252.25 mmol), phthalimide (66.8 g, 454.02 mmol) and triphenylphosphine(119 g, 453.70 mmol) in toluene/THF (200/200 mL) was cooled undernitrogen with ice. DIAD (91.8 g, 453.98 mmol) was added dropwise over 20minutes and the mixture was stirred at 0° C. for 20 minutes, withoutcooling for 2 hr. The mixture was filtered and the filtrate wasevaporated. The residue was taken up in ether and filtered. The filtratewas evaporated and the residue was purified by silica gel column andeluted with EtOAc/hexane (0-50%) to give the title compound (74 g, 79%)as a colorless oil.

MS: 323 ES+ (C₂₄H₂₆N₂O₅)

Intermediate 156: (R)-tert-butyl1-aminobut-3-en-2-yl(benzyloxy)carbamate

A solution of (R)-tert-butylbenzyloxy(1-(1,3-dioxoisoindolin-2-yl)but-3-en-2-yl)carbamate(Intermediate 155, 80 g, 189.36 mmol) and hydrazine hydrate (47.4 g,946.86 mmol) in MeOH (800 mL) was stirred at rt overnight. The mixturewas diluted with ether and filtered. The filtrate was evaporated and theresidue was purified by silica gel column and eluted with MeOH/DCM(0-10%) to give the title compound (44.3 g, 80%) as a colorless oil.

¹H NMR (300 MHz, METHANOL-d) δ: 7.36-7.40 (m, 5H), 5.76-5.88 (m, 1H),5.14-5.21 (m, 2H), 4.83 (q, 2H), 4.27-4.30 (m, 1H), 2.81-2.88 (m, 1H),2.67-2.73 (m, 1H), 1.45 (s, 9H).

Intermediate 157: 2-(((tert-butyldimethylsilyl)oxy)methyl)prop-2-en-1-ol

To a dry flask containing sodium hydride (2.270 g, 56.75 mmol) in THF(150 mL) under N₂ at 0° C. was added slowly 2-methylenepropane-1,3-diol(5 g, 56.75 mmol). The reaction was warmed to room temperature andstirred 45 min. TBS-Cl (8.55 g, 56.75 mmol) was added in one batch andstirring was continued for another 45 min, until complete by TLC. To thereaction was added water and the mixture was extracted three times withEtOAc and washed with brine. The organics were dried over magnesiumsulfate, filtered and concentrated at −10-15° C. Silica gelchromatography (20%-50% EtOAc/Hex) afforded the title compound (11.36 g,99%) as a clear oil.

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (s, 6H) 0.88 (s, 10H) 3.91 (d, J=5.65Hz, 2H) 4.10 (s, 2H) 4.74 (t, J=5.46 Hz, 1H) 5.00 (t, J=1.51 Hz, 2H).

Intermediate 158: 2-(((tert-butyldimethylsilyl)oxy)methyl)acrylaldehyde

To a solution of 2-(((tert-butyldimethylsilyl)oxy)methyl)prop-2-en-1-ol(Intermediate 157, 11.35 g, 56.09 mmol) in DCM (150 mL) was addedmanganese dioxide (activated, 5 g) (28.7 g, 280.43 mmol). The reactionmixture was stirred at room temperature for three days. More MnO₂ wasadded and continued stirring until TLC shows minimum starting material.The reaction mixture was filtered through a pad of silica, evaporated at˜15° C. and purified on silica gel column (5-20%, EA/Hex) to afford thetitle compound (6.70 g, 59.6%) as a colorless oil.

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.06-0.13 (m, 6H) 0.89-0.97 (m, 9H)4.41 (t, J=2.07 Hz, 2H) 6.11 (q, J=1.88 Hz, 1H) 6.50-6.56 (m, 1H) 9.63(s, 1H).

Intermediate 159: tert-butyl((2R)-1-((((9H-fluoren-9-yl)methoxy)carbonyl)(2-(((tert-butyldimethylsilyl)oxy)methyl)-1-cyanoallyl)amino)but-3-en-2-yl)(benzyloxy)carbamate

2-(((tert-butyldimethylsilyl)oxy)methyl)acrylaldehyde (Intermediate 158,4.76 g, 23.76 mmol) was added to a solution of (R)-tert-butyl(1-aminobut-3-en-2-yl)(benzyloxy)carbamate (Intermediate 156, 5.79 g,19.80 mmol) in THF (100 mL), followed by addition of trimethylsilylcyanide (10.62 mL, 79.19 mmol). The mixture was stirred at roomtemperature overnight. More TMS-CN was added and stirred longer untilless starting material seen by LCMS. MgSO₄ was added to the reactionmixture and stirred for 1 hr. Sodium bicarbonate (3.33 g, 39.58 mmol)was added, followed by (9H-fluoren-9-yl)methyl carbonochloridate (7.68g, 29.69 mmol). The resulting mixture was stirred at room temperatureovernight. 0.3 eq more FmocCl was added and stirred 5 h more. Themixture was filtered and the filtrate was evaporated. The residue waspurified on silica gel column (0-22%, EtOAc/hexane) to afford the titlecompound (3.32 g, 23.18%) as a pale yellow thick oil.

MS: 724 ES+ (C₄₂H₅₃N₃O₆Si)

Intermediate 160: (5R)-(9H-fluoren-9-yl)methyl5-((benzyloxy)(tert-butoxycarbonyl)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-cyano-5,6-dihydropyridine-1(2H)-carboxylate

A solution of tert-butyl((2R)-1-((((9H-fluoren-9-yl)methoxy)carbonyl)(2-(((tert-butyldimethylsilyl)oxy)methyl)-1-cyanoallyl)amino)but-3-en-2-yl)(benzyloxy)carbamate(Intermediate 159, 3.32 g, 4.59 mmol) in toluene (150 mL) was bubbledwith nitrogen for 15 minutes. Hoveyda-Grubbs Catalyst 2nd Generation(0.288 g, 0.46 mmol) was added and the mixture was bubbled with nitrogenfor an additional 15 minutes. The resulting solution was heated undernitrogen at 90° C. for 2 days. The reaction mixture was evaporated. Theresidue was purified on silica gel column (0-30% EtOAc/hexane) to affordthe title compound (1.300 g, 40.7%) as a light brown film.

MS: 596 ES+ (C₄₀H₄₉N₃O₆Si)

Intermediate 161: (5R)-(9H-fluoren-9-yl)methyl5-((benzyloxy)(tert-butoxycarbonyl)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate

Preparation of copper(II) chloride on 4 Å molecular sieves: 4 ÅMolecular sieves (2 g, 2.00 mmol) was added to a solution of copper(II)chloride dihydrate (0.341 g, 2.00 mmol) in water (200 mL) and thesuspension was stirred overnight. The solid was filtered, washed withwater and acetone, and dried in an oven at 140° C. for 1 hr to obtain2.104 g of blue solid. The material needed to be activated by puttinginto 140° C. over for a couple hours and cooled to room temperature in adesicator prior to use.

A mixture of (5R)-(9H-fluoren-9-yl)methyl5-((benzyloxy)(tert-butoxycarbonyl)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-cyano-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 160, 1.3 g, 1.87 mmol), acetaldehyde oxime (0.569 mL, 9.34mmol) and newly activated copper(II) chloride on 4 Å MS (187 mg, 0.15mmol) (used 100 mg CuCl₂/mol sieves per 1 mmol substrate) in MeOH (5 mL)was stirred under nitrogen at 65° C. for 6 h. The mixture was filteredand evaporated. The residue was purified on silica gel column (0-60%,EtOAc/hexanes). The two diastereomers were combined together to affordthe title compound (1.190 g, 89%) as a pale yellow foamy solid.

MS: 714 ES+ (C₄₀H₅₁N₃O₇Si)

Intermediate 162: (5R)-(9H-fluoren-9-yl)methyl5-((benzyloxy)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (5R)-(9H-fluoren-9-yl)methyl5-((benzyloxy)(tert-butoxycarbonyl)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 161, 1.19 g, 1.67 mmol) in DCM (20 mL) at room temperaturewas added zinc bromide (1.502 g, 6.67 mmol). The reaction mixture wasstirred at room temperature for 2 h. The reaction mixture was dilutedwith dichloromethane and washed with saturated sodium bicarbonate and.The aqueous wash was extracted three to four times more with DCM. Theorganics were washed with brine, dried over magnesium sulfate, filteredand concentrated to afford the title compound (0.970 g, 95%) as an offwhite foam.

MS: 614 ES+ (C₃₅H₄₃N₃O₅Si)

Intermediate 163:(5R)-5-((benzyloxy)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,5,6-tetrahydropyridine-2-carboxamide

Diethylamine (0.824 mL, 7.89 mmol) was added to a solution of(5R)-(9H-fluoren-9-yl)methyl5-((benzyloxy)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 162, 0.968 g, 1.58 mmol) in DCM (20 mL) and the resultingsolution was allowed to stir at room temperature overnight. Thevolatiles were removed by evaporation and the residue was purified onsilica gel (0-12%, MeOH/DCM, UV 220 nm) to afford the title compound(0.564 g, 91%) as a yellow film.

MS: 392 ES+ (C₂₀H₃₃N₃O₃Si)

Intermediate 164:(2S,5R)-6-(benzyloxy)-3-(((tert-butyldimethylsilyl)oxy)methyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of(5R)-5-((benzyloxy)amino)-3-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 163, 0.562 g, 1.44 mmol) and diisopropylethyl amine (1.000mL, 5.74 mmol) in acetonitrile (200 mL) at 0° C. was added triphosgene(0.145 g, 0.49 mmol) as a solution in acetonitrile (6 mL). Thetriphosgene solution was added at a rate of 4 mL/hour. Once addition wascomplete the reaction was stirred at 0° C. overnight. The reactionmixture was diluted with ethyl acetate and washed with saturated sodiumbicarbonate. The aqueous layer was extracted again with EtOAc and thenDCM. The combined organic extracts were washed with brine, dried overmagnesium sulfate, filtered and concentrated. Silica gel chromatography(0%-100, ethyl acetate/hexanes) afforded the title compound (0.225 g,37.5%) as a colorless film.

MS: 418 ES+ (C₂₁H₃₁N₃O₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: −0.08 (s, 6H) 0.76 (s, 9H) 2.89-3.00(m, 1H) 3.04-3.13 (m, 1H) 3.32 (dd, J=5.09, 2.45 Hz, 1H) 4.01-4.09 (m,1H) 4.19-4.28 (m, 1H) 4.30 (s, 1H) 4.67-4.76 (m, 1H) 4.83-4.93 (m, 1H)5.51 (br. s., 1H) 6.09-6.21 (m, 1H) 6.63 (br. s., 1H) 7.19-7.36 (m, 5H).

Intermediate 165: trimethylammonium(2S,5R)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

To a solution of(2S,5R)-6-(benzyloxy)-3-(((tert-butyldimethylsilyl)oxy)methyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 164, 30 mg, 0.07 mmol) in EtOAc (1.33 mL), water (1.995mL) and EtOH (0.660 mL) was added Pd/C (wet, degussa type E101 NE/W)(7.65 mg, 7.18 μmol), SO₃.TMA (12.00 mg, 0.09 mmol) and TEA (2.003 μl,0.01 mmol) under N₂ atmosphere. The reaction mixture was degassed andfilled with H₂ using a balloon. The reaction mixture was stirred at roomtemperature under a H₂ balloon for 5 hours. The H₂ balloon was removedand the reaction mixture was stirred overnight. The reaction mixture wasfiltered and the organics evaporated. The remaining aqueous mixture waslyophilized and the resulting residue was purified by preparative HPLC(Synergi Polar RP 21.2 mm×100 mm 4 μm coupled with YMC C30 20 mm×100 mm5 μm) to afford the title compound as a white solid, 5 mg, 15%.

MS: 406 ES− (C₁₄H₂₄N₃O₇SSi)

Intermediate 166:(2S,5R)-2-carbamoyl-3-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

Dowex 50WX8, 100-200 mesh (0.75 g, 0.01 mmol) was conditioned bystirring for 3 hours in 2N NaOH (1.8 mL, 3.60 mmol). The resin was thenloaded into a cartridge and washed with water until the pH was 7. It wasthen washed with (1/1) acetone/water, followed by water.(2S,5R)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 165, 5 mg, 0.01 mmol) in water was passed throughthe resin 5 times and then lyophilized to afford the title compound(3.00 mg, 56.7%).

MS: 406 ES+ (C₁₄H₂₄N₃O₇SSi)

Intermediate 167:(2S,5R)-2-carbamoyl-3-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate

Dowex 50WX8, 100-200 mesh (0.48 g, 6.43 μmol) resin in 2 mL water wasloaded into a cartridge and let water run off. Trimethylammonium(2S,5R)-3-(((tert-butyldimethylsilyl)oxy)methyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate sodium salt (Intermediate 166, 3 mg, 6.43 μmol) in water waspassed through the resin 2 times (pH is ˜3). The solution was carriedforward to the final ion exchange column, see Example 18.

Example 19(2S,5R)-4-(2-amino-2-oxoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(2S,5R)-2-carbamoyl-4-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 179, 34 mg, 0.04 mmol) according to the proceduredescribed for Example 1. It was further purified on reverse phase HPLCusing Synergi Polar RP (21.2 mm×100 mm 4 μm) coupled with YMC C30(20×150 mm 5 μm) (Mobile Phase A 100% H₂O, Mobile Phase B 100%Acetonitrile), yielding a pale yellow solid, 1.6 mg, 3%.

MS: 343 ES+ (C₉H₁₂N₄NaO₇S)

¹H NMR (300 MHz, D₂O) δ: 3.17 (s, 2H); 3.37 (d, 1H); 3.70 (dd, 1H); 4.23(d, 1H); 4.64 (d, 1H); 5.93 (d, 1H).

The Intermediates for Example 19 were Prepared as Follows Intermediate168: (S)-tert-butyl2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(((4-methoxybenzyl)-oxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (2S)-tert-butyl5-hydroxy-4-(hydroxymethyl)-2-(isopropyldimethylsilyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 117, 13.0 g, 34.99 mmol) in toluene (150 mL) was added4-methoxybenzyl 2,2,2-trichloroacetimidate (7.65 mL, 10.42 g, 36.74mmol) and La(OTf)₃ (205 mg, 0.35 mmol). The mixture was stirred at 50°C. for 4 h. Aqueous work-up with ethyl acetate and the organic layer wasdried over MgSO₄ to afford the crude product.

MS: 492 ES+ (C₂₆H₄₁NO₆Si)

Intermediate 169: (2S)-tert-butyl2-(((tert-butyldimethylsilyl)oxy)methyl)-5-hydroxy-4-(((4-methoxybenzyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(((4-methoxybenzyl)oxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 168, 34.99 mmol) according to the procedure forIntermediate 8. The reaction mixture was concentrated and the whitesolid was redissolved in 200 mL EtOAc and washed with satd. NaHCO₃,brine, dried over MgSO₄, filtered and concentrated. The residue waspurified by silica gel column eluting with 0-100% ethyl acetate/hexanesto afford a colorless oil. 17.10 g, 99% over 2 steps.

MS: 494 ES+ (C₂₆H₄₃NO₆Si)

¹H NMR (300 MHz, CDCl₃) δ: 0.01 (s, 6H); 0.83 (s, 9H); 1.41 (s, 9H);3.62 (m, 1H); 3.75 (s, 3H); 3.75 (m, 1H); 4.14 (m, 4H); 4.34 (m, 1H);4.41 (s, 2H); 5.70 (d, 1H); 6.83 (d, 2H); 7.21 (d, 2H).

Intermediate 170: (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(((tert-butyldimethylsilyl)-oxy)methyl)-4-(((4-methoxybenzyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(((4-methoxybenzyl)-oxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 169, 17.10 g, 34.78 mmol) andN-(allyloxy)-2-nitrobenzenesulfonamide (Intermediate 9, 9.88 g, 38.26mmol) according to the procedure described for Intermediate 10. Desiredproduct was obtained as a pale yellow oil, 18.70 g, 74%.

MS: 734 ES+ (C₃₅H₅₁N₃O₁₀SSi)

¹H NMR (300 MHz, CDCl₃) δ: 0.05 (s, 6H); 0.87 (s, 9H); 1.26 (s, 9H);3.71 (m, 2H); 3.81 (s, 3H); 4.45 (m, 4H); 4.57 (d, 2H); 5.28 (m, 2H);5.32 (m, 2H); 5.71 (m, 1H); 5.93 (m, 2H); 6.12 (d, 1H); 6.86 (m, 2H);7.70 (m, 5H); 8.13 (m, 1H).

Intermediate 171: (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(((4-methoxybenzyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 170, 18.70 g, 25.48 mmol) in DCM/water (180 mL/20 mL), wasadded DDQ (6.94 g, 30.57 mmol) at 0° C. The mixture was stirred at roomtemperature for 1 hour before it was concentrated. The residue waspurified on silica gel column eluting with 0-100% ethyl acetate/hexanesto give a pale yellow oil, 12.38 g, 79%.

MS: 614 ES+ (C₂₇H₄₃N₃O₉SSi)

¹H NMR (300 MHz, CDCl₃) δ: 0.05 (s, 6H); 0.87 (s, 9H); 1.40 (s, 9H);1.81 (bs, 1H); 3.63 (d, 1H); 3.70 (m, 2H); 4.27 (m, 6H); 4.60 (m, 1H);5.18 (m, 2H); 5.71 (m, 1H); 6.09 (m, 1H); 7.60 (t, 1H); 7.77 (m, 2H);8.14 (t, 1H).

Intermediate 172: (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(cyanomethyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred solution of (2S)-tert-butyl5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 171, 12.38 g, 20.17 mmol) in DCM (200 mL) at 0° C.,pyridine (2.39 g, 30.25 mmol) and N,N-dimethylpyridin-4-amine (123 mg,1.01 mmol) was added. Then methanesulfonic anhydride (4.22 g, 24.20mmol) was added. The mixture was then stirred from 0° C. to roomtemperature for 2 hours. It was diluted with DCM (100 mL) and washedwith brine, dried over MgSO₄, filtered and concentrated to give thecrude as a pale yellow oil. It was used directly for the next step.

To a stirred solution of sodium cyanide (4.94 g, 100.85 mmol) in water(20.0 mL) was added (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(((methylsulfonyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate(crude, 20.17 mmol) in DMF (100 mL). The mixture was then stirred atroom temperature for 15 h. It became a dark orange solution. It was thendiluted with sat. NaHCO₃ and water and extracted with ethyl acetate,dried over MgSO₄. The crude was subjected to silica gel column elutingwith 0-100% ethyl acetate/hexanes to give a pale yellow oil, 5.92 g,47.1%.

MS: 623 ES+ (C₂₈H₄₂N₄O₈SSi)

¹H NMR (300 MHz, CDCl₃) δ: 0.05 (s, 6H); 0.88 (s, 9H); 1.41 (s, 9H);3.12 (m, 1H); 3.32 (m, 1H); 3.74 (m, 2H); 4.21 (m, 5H); 4.65 (s, 1H);5.18 (m, 2H); 5.69 (m, 1H); 6.31 (bs, 1H); 7.65 (m, 1H); 7.81 (m, 2H);8.15 (d, 1H).

Intermediate 173: (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate

Preparation of the catalyst: 4 Å molecular sieves (MS-4 Å) wereimpregnated with the corresponding metal salt (CuCl₂-2H₂O) as follows: 1mmol of the salts was dissolved in 100 ml of deionized H₂O and stirredwith 1 g of MS-4 Å at room temperature for 6 h. The solid was filtered,washed with deionized H₂O and acetone, and dried in an oven at 150° C.for 1 h. (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(cyanomethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 172, 3.60 g, 5.78 mmol), Cu(II)Cl₂-4 Å catalyst (200 mg),acetaldoxime (1.762 mL, 28.90 mmol, 5.0 eq) and MeOH (40.0 mL) werestirred at 60° C. for 15 h. The solid was filtered, washed with MeOH andthe filtrate was evaporated to afford crude product.

MS: 641 ES+ (C₂₈H₄₂N₄O₈SSi)

Intermediate 174: (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 173, crude, 5.78 mmol) following the procedure describedfor Intermediate 18. Silica column eluting with 0-100% ethylacetate/hexanes gave a pale yellow solid. 2.61 g, 86%.

MS: 527 ES+ (C₂₂H₃₀N₄O₉S)

¹H NMR (300 MHz, CDCl₃) δ: 1.39 (s, 9H); 3.03 (m, 2H); 3.33 (m, 2H);3.70 (m, 2H); 4.20 (m, 1H); 4.35 (m, 2H); 4.68 (bs, 1H); 5.17 (m, 2H);5.72 (m, 2H); 6.00 (s, 1H); 6.08 (bs, 1H); 6.32 (bs, 1H); 7.65 (d, 1H);7.80 (m, 2H); 8.15 (d, 1H).

Intermediate 175: (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate

Stock solution: 240 mg conc. HNO₃ and 80 mg Na₂Cr₂O₇.2H₂O was dissolvedin 16 mL H₂O at rt.

To the solution of (25)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 174, 2.61 g, 4.96 mmol) in acetonitrile (50 mL) was addedsodium periodate (4.66 g, 21.81 mmol) and catalytic solution ofNaCr₂O₇/HNO₃ solution (6.0 mL) at room temperature and stirred for 15 h.The mixture was diluted with 100 mL ethyl acetate, 25 mL 1M pH 7 buffer,25 mL 2M NaHSO₃. The aqueous solution was extracted with 50 mL ethylacetate. The ethyl acetate layer was then washed with brine and driedover MgSO₄, filtered and concentrated to afford(2S)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-1-(tert-butoxycarbonyl)-1,2,5,6-tetrahydropyridine-2-carboxylicacid, which was used directly in the next step without furtherpurification. Pale yellow solid, crude, 2.79 g, 104%.

MS: 541 ES+ (C₂₂H₂₈N₄O₁₀S)

To a stirred solution of(2S)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-1-(tert-butoxycarbonyl)-1,2,5,6-tetrahydropyridine-2-carboxylicacid (crude, 2.79 g, 5.16 mmol), ammonium chloride (552 mg, 10.32 mmol),HATU (3.93 g, 10.32 mmol) in DMF (20 mL) at 0° C., was added DIPEA (2.67g, 20.65 mmol). After stirring at room temperature for 1 hr, 50 mL EtOAcand 50 mL water was added. The organic layer was washed with water,brine and dried over MgSO₄, filtered and concentrated to give a residue,which was purified on silica gel column eluting with 0-100% ethylacetate/hexanes to afford an off-white solid, 1.36 g, 49%.

MS: 540 ES+ (C₂₂H₂₉N₅O₉S)

¹H NMR (300 MHz, CDCl₃) δ: 1.38 (s, 9H); 3.08 (m, 2H); 4.24 (m, 1H);4.40 (m, 2H); 5.17 (m, 2H); 5.68 (m, 1H); 6.02 (m, 2H); 6.20 (m, 2H);6.40 (bs, 1H); 6.51 (bs, 1H); 7.64 (d, 1H); 7.78 (m, 2H); 8.14 (d, 1H).

Intermediate 176: (2S)-tert-butyl5-((allyloxy)amino)-4-(2-amino-2-oxoethyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (2S)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-(2-amino-2-oxoethyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 175, 1.36 g, 2.52 mmol) and potassium carbonate (1.742 g,12.60 mmol) in acetonitrile (20.0 mL) at 0° C. was added benzenethiol(1.389 g, 12.60 mmol). The mixture was stirred at room temperature for15 hours. The reaction mixture was concentrated and the resultingresidue was triturated with DCM. The solids were removed by filtration.The filtrate was concentrated to a pale yellow film, which was subjectedto silica gel column eluting with 0-50% MeOH/DCM to give a pale yellowsolid, 330 mg, 36.9%.

MS: 355 ES+ (C₁₆H₂₆N₄O₅)

¹H NMR (300 MHz, CDCl₃) δ: 1.50 (s, 9H); 3.05 (m, 2H); 3.20 (m, 1H);3.45 (bs, 1H); 4.23 (d, 2H); 4.500 (m, 1H); 5.03 (m, 1H); 5.24 (m, 2H);5.88 (m, 2H); 5.94 (m, 2H); 6.57 (bs, 1H); 6.76 (bs, 1H).

Intermediate 177:(R)-5-(allyloxyamino)-4-(2-amino-2-oxoethyl)-1,2,5,6-tetrahydropyridine-2-carboxamide

To a solution of (2S)-tert-butyl5-((allyloxy)amino)-4-(2-amino-2-oxoethyl)-2-carbamoyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 176, crude, 330 mg, 0.93 mmol) in DCM (5.0 mL) at 0° C.was added HCl (4.0M in dioxane, 0.233 mL, 0.93 mmol). The mixture wasstirred from 0° C. to room temperature for 2 h. Then 0.233 mL more HClwas added to the mixture and stirred for 2 h. It was neutralized byK₂CO₃ and TEA, filtered and the solvent was removed under vacuum. Thecrude was subjected to silica column eluting with 0-50% MeOH/DCM to givethe desired diastereomeric products as white solids. 100 mg,diastereomer 1. 32 mg, diastereomer 2. 56% yield.

Diastereomer 1:

MS: 255 ES+ (C₁₁H₁₈N₄O₃)

¹H NMR (300 MHz, CD₃OD) δ: 3.22 (m, 1H); 3.38 (m, 2H); 3.61 (m, 2H);3.82 (m, 1H); 4.27 (d, 2H); 4.75 (m, 1H); 5.30 (m, 2H); 6.03 (m, 1H);6.15 (d, 1H).

Diastereomer 2:

MS: 255 ES+ (C₁₁H₁₈N₄O₃)

¹H NMR (300 MHz, CD₃OD) δ: 3.16 (m, 2H); 3.33 (m, 2H); 3.58 (m, 2H);4.16 (m, 2H); 4.38 (bs, 1H); 5.19 (m, 2H); 5.89 (d, 1H); 5.95 (m, 1H).

Intermediate 178:(2S,5R)-6-(allyloxy)-4-(2-amino-2-oxoethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

The title compound was prepared from(R)-5-(allyloxyamino)-4-(2-amino-2-oxoethyl)-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 177, 132 mg, 0.52 mmol) following the procedure describedfor Intermediate 16. The reaction mixture was stirred at 0° C. for 2 h.The volatile was removed under vacuum and the crude was subjected tosilica column eluting with 0-100% ethyl acetate/hexanes and then 95%ethyl acetate/5% MeOH to give a pale yellow solid. 62 mg, 43%.

MS: 281 ES+ (C₁₂H₁₆N₄O₄)

¹H NMR (300 MHz, CD₃) δ: 3.13 (m, 3H); 3.43 (dd, 1H); 4.10 (d, 1H); 4.43(m, 3H); 5.35 (m, 2H); 5.57 (bs, 1H); 5.80 (m, 2H); 5.90 (d, 1H); 6.01(m, 1H); 6.86 (bs, 1H).

Intermediate 179:(2S,5R)-4-(2-amino-2-oxoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-4-(2-amino-2-oxoethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 178, 62 mg, 0.22 mmol) following the procedure describedfor Intermediate 17. The desired product was obtained as a pale yellowoil. 34 mg, 48%.

MS: 304; 320 (C₂₁H₂OP; C₉H₁₁N₄O₇S).

¹H NMR (300 MHz, CD₃Cl) δ: 2.31 (m, 1H); 2.64 (s, 3H); 3.15 (m, 4H);3.63 (dd, 1H); 4.22 (d, 1H); 4.45 (bs, 1H); 5.62 (bs, 1H); 5.96 (d, 1H);6.01 (bs, 1H); 6.64 (bs, 1H); 6.96 (bs, 1H); 7.68 (m, 15H); 8.60 (bs,1H).

Example 20(2S,5R)-4-carbamoyl-2-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The DOWEX resin 50WX8 (50 g) was conditioned by stirring for 20 min in2N sodium hydroxide (40 ml). The DOWEX was loaded into a cartridge,washed with water until pH=7, then washed with acetone/water (1/1) andwater again. The compound (Intermediate 180, 0.31 g, 0.76 mmol) wasloaded using water and a minimum amount of acetone and eluted withwater. The fractions were reloaded onto the cartridge and washed thoughwith water again. This load and wash process was repeated five times,then the final round of fractions was directly frozen and lyophilized togive a white solid (130 mg). Reversed phase chromatography (0%-4% MeCNin water) afforded the title compound as a white solid (31 mg).

MS: 292 ES− (C₈H₁₀N₃O₇S)

¹H NMR (600 MHz, DEUTERIUM OXIDE) δ: 3.56 (d, J=11.67 Hz, 1H); 3.64 (br.s., 1H); 3.69-3.81 (m, 1H); 3.97 (d, J=6.40 Hz, 2H); 4.18 (br. s., 1H);4.84 (br. s., 1H); 6.73 (br. s., 1H).

The Intermediates for Example 20 were Prepared as Follows Intermediate180:(2S,5R)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate-(E)-triphenyl(prop-1-en-1-yl)phosphonium

To a solution of(2S,5R)-6-(allyloxy)-2-(((tert-butyldimethylsilyl)oxy)methyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide(Intermediate 108, 0.32 g, 0.87 mmol) in DCM (15 mL) was added aceticacid (dried over sodium sulfate) (0.100 mL, 1.74 mmol) andtetrakis(triphenylphosphine)palladium(0) (1.006 g, 0.87 mmol) undernitrogen at room temperature. The yellow solution was stirred at roomtemperature for 3.5 hours. To the mixture was added pyridine (10 ml),followed by sulfur trioxide pyridine complex (0.832 g, 5.22 mmol). Thesuspension was stirred at room temperature overnight under nitrogen. Thereaction mixture was concentrated to dryness (≦30° C.), suspended inDCM, filtered, concentrated and purified on a silica gel column (0%-100%MeOH in DCM) to afford the title compound as a white solid (0.31 g).

MS: 406 ES− (C₁₄H₂₄N₃O₇SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.07-0.05 (m, 6H); 0.71-0.86 (m, 9H); 2.10(dt, J=6.28, 2.05 Hz, 3H); 3.03-3.18 (m, 1H); 3.22-3.32 (m, 1H);3.60-3.73 (m, 1H); 3.79 (d, J=6.42 Hz, 2H); 4.48 (d, J=3.02 Hz, 1H);6.40 (dd, J=2.93, 1.04 Hz, 1H); 6.43-6.70 (m, 1H); 7.17 (br. s., 2H);7.56-7.96 (m, 16H).

Example 21(2S,5R)-2-carbamoyl-3,4-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3,4-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 199, 0.22 g, 0.37 mmol) following the proceduredescribed for Example 1. The desired product was obtained as a whitesolid (99.6 mg, 86%).

Optical rotation: (0.2 g/dL, MeOH)=−228

MS: 290.0 ES− (C₉H₁₃N₃O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.51 (s, 3H); 1.75 (s, 3H); 3.03 (dd,J=10.76, 3.02 Hz, 1H); 3.70 (d, J=10.76 Hz, 1H); 3.88-4.06 (m, 2H); 7.23(br. s., 1H); 7.76 (br. s., 1H).

The Intermediates for Example 21 were Prepared as Follows Intermediate181: (R)-3-tert-butyl 4-methyl 2,2-dimethyloxazolidine-3,4-dicarboxylate

A solution of (R)-methyl2-(tert-butoxycarbonylamino)-3-hydroxypropanoate (Aldrich, 9.26 mL,45.61 mmol) in dichloromethane (18.10 mL) was cooled to 0° C. in anice-water bath. After 20 minutes 2,2-dimethoxypropane (Aldrich, 28.6 mL,232.63 mmol) and para-toluene sulfonic acid monohydrate (1.301 g, 6.84mmol) were added at 0° C. and stirred overnight at 25° C. After 24 hoursobserved starting material still present, added additional 270 mg ofpara-toluene sulfonic acid monohydrate and let reaction mixture stir at25° C. for 3 hours. The reaction mixture was poured into saturatedaqueous NaHCO₃ solution and the resulting solution was extracted withether. The combined organic layers were washed with sodium bicarbonatesolution and brine. The resulting organic layers were dried overmagnesium sulfate, filtered, and concentrated. Silica gel chromatography(0%-70% ethyl acetate/hexanes) afforded a colorless oily solid (9.53 g,81%).

MS: 260.1 ES+ (C₁₂H₂₁NO₅)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.37-1.57 (m, 12H); 1.62-1.71 (m, 3H);3.70-3.81 (m, 3H); 3.98-4.22 (m, 2H); 4.33-4.54 (m, 1H).

Intermediate 182: (R)-tert-butyl4-formyl-2,2-dimethyloxazolidine-3-carboxylate

To a −78° C. solution of (R)-3-tert-butyl 4-methyl2,2-dimethyloxazolidine-3,4-dicarboxylate (Intermediate 181, 18.80 g,72.50 mmol) in toluene (153 mL) was dropwise (via addition funnel) addedDIBAL-H (1.0 M in toluene) (116 ml, 116.01 mmol) over 1.5 hours. Thereaction mixture was stirred at −78° C. for 2 hours under nitrogenatmosphere. The cold reaction mixture was treated with 38 mL MeOH (˜1.91M) and then poured 750 mL cold 1N HCl (˜0.09M) into reaction mixture,removed reaction mixture from −78° C. bath, transferred to separatoryfunnel and extracted with ethyl acetate. The organic layers were driedover magnesium sulfate, filtered, concentrated. Due to a difficultseparation required two silica gel chromatography columns (0%-50% ethylacetate/hexanes) to afford the title compound as a colorless oily solid(10.2 g, 61%).

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.36-1.74 (m, 15H); 3.95-4.49 (m, 3H);9.45-9.68 (m, 1H).

Intermediate 183: (R)-tert-butyl4-(1-hydroxyethyl)-2,2-dimethyloxazolidine-3-carboxylate

To a solution of (R)-tert-butyl4-formyl-2,2-dimethyloxazolidine-3-carboxylate (Intermediate 182, 10.18g, 44.40 mmol) in THF (188 mL) at −78° C. was added methyl magnesiumbromide (16.28 mL, 48.84 mmol) dropwise. The reaction mixture wasallowed to warm to room temperature and stir overnight under nitrogenatmosphere. Re-cooled reaction mixture to −78° C. and added dropwiseadditional 13 mL (0.9 eq, to bring total to 2 eq) of methyl magnesiumbromide. Allowed reaction mixture to stir at −78° C. for 3 hours andwarm to room temperature. The reaction mixture was quenched with waterand diluted with ethyl acetate saturated sodium chloride (brine). Theresulting emulsion was filtered thru celite and the layers separated.The organic layers were dried over magnesium sulfate, filtered,concentrated. Silica gel chromatography (0%-20% ethyl acetate/hexanes)afforded the title compound as a colorless oily solid (8.42 g, 77%).

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.09-1.22 (m, 3H); 1.36-1.64 (m, 15H);3.70-4.27 (m, 4H).

Intermediate 184: (R)-tert-butyl4-acetyl-2,2-dimethyloxazolidine-3-carboxylate

The title compound was prepared from (R)-tert-butyl4-(1-hydroxyethyl)-2,2-dimethyloxazolidine-3-carboxylate (Intermediate183, 8.42 g, 34.32 mmol) following the procedure for Intermediate 231.Silica gel chromatography (0%-50% ethyl acetate/hexanes) afforded thedesired product as a colorless oily solid (8.16 g, 98%).

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.37-1.57 (m, 12H); 1.61-1.77 (m, 3H);2.20 (br. s., 3H); 3.89-4.03 (m, 1H); 4.07-4.21 (m, 1H); 4.24-4.47 (m,1H).

Intermediate 185: (S)-tert-butyl2,2-dimethyl-4-(prop-1-en-2-yl)oxazolidine-3-carboxylate

The title compound was prepared from (R)-tert-butyl4-acetyl-2,2-dimethyloxazolidine-3-carboxylate (Intermediate 184, 8.16g, 33.54 mmol) following the procedure for JC75. Silica gelchromatography (0%-20% ethyl acetate/hexanes) afforded the desiredproduct as a colorless oily solid (7.25 g, 90%).

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.37-1.54 (m, 12H); 1.66 (br. s., 3H);1.74 (s, 3H); 3.75 (dd, J=9.06, 3.02 Hz, 1H); 4.02-4.19 (m, 1H);4.20-4.44 (m, 1H); 4.86 (br. s., 2H).

Intermediate 186: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-ylcarbamate

The title compound was prepared from (S)-tert-butyl2,2-dimethyl-4-(prop-1-en-2-yl)oxazolidine-3-carboxylate (Intermediate185, 6.92 g, 28.67 mmol) following the procedure for JC76. Silica gelchromatography (0%-10% ethyl acetate/hexanes) afforded the desiredproduct as a colorless oil (8.28 g, 92%).

MS: 316.3 ES+ (C₁₆H₃₃NO₃Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.05 (d, J=2.27 Hz, 6H); 0.84-0.93 (m,9H); 1.45 (s, 9H); 1.76 (s, 3H); 3.59-3.81 (m, 2H); 4.06 (d, J=6.99 Hz,1H); 4.81-5.05 (m, 2H).

Intermediate 187:(S)-1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-amine

The title compound was prepared from (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-ylcarbamate(Intermediate 186, 8.28 g, 26.24 mmol) following the procedure forIntermediate 234. The organic layers were dried over magnesium sulfate,filtered and concentrated to afford the title compound as an off-whitesolid (5.57 g, 99%).

MS: 216.2 ES+ (C₁₁H₂₅NOSi)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.06-0.15 (m, 6H); 0.88-0.96 (m, 9H);1.78-1.88 (m, 3H); 3.00 (br. s., 2H); 3.53-3.64 (m, 1H); 3.66-3.79 (m,1H); 3.91 (dd, J=10.29, 3.30 Hz, 1H); 4.97-5.11 (m, 2H).

Intermediate 188: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate

The title compound was prepared from(S)-1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-amine(Intermediate 187, 5.57 g, 25.86 mmol) and2-bromo-N-methoxy-N-methylacetamide (Intermediate 4, 4.28 g, 23.53 mmol)following the procedure described for Intermediate 5. Silica gelchromatography (0%-20% ethyl acetate/hexanes) afforded the desiredproduct as a colorless oil (4.24 g, 39%).

MS: 417.3 ES+ (C₂₀H₄₀N₂O₅Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: −0.03-0.15 (m, 6H); 0.81-0.97 (m, 9H);1.41-1.63 (m, 9H); 1.72-1.82 (m, 3H); 3.55-3.79 (m, 2H); 3.92-4.23 (m,1H); 4.85-5.02 (m, 2H).

Intermediate 189: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-yl(3-methyl-2-oxobut-3-enyl)carbamate

The title compound was prepared from (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 188, 4.24 g, 10.18 mmol) following the procedure describedfor Intermediate 6 substituting the relevant isoprenyl magnesium bromide(0.5 M in THF) (204 mL, 101.77 mmol). Silica gel chromatography (0%-20%ethyl acetate/hexanes) afforded the desired product as a light yellowoil (3.72 g, 92%).

MS: 398.3 ES+ (C₂₁H₃₉NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: −0.02-0.08 (m, 6H); 0.82-0.90 (m, 9H);1.35-1.51 (m, 9H); 1.78 (s, 3H); 1.90 (s, 3H); 3.68-3.99 (m, 2H);4.23-4.75 (m, 3H); 4.84-5.01 (m, 2H); 5.84-5.98 (m, 2H).

Intermediate 190: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3,4-dimethyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-methylbut-3-en-2-yl(3-methyl-2-oxobut-3-enyl)carbamate(Intermediate 189, 3.72 g, 9.36 mmol) following the procedure describedfor Intermediate 7 except the reaction mixture was heated at 110-120° C.for 48 hours and doubling the catalyst loading (0.6 equivalents). Silicagel chromatography (0%-15% ethyl acetate/hexanes) afforded the desiredproduct as a light tan oily solid (2.39 g, 69%).

Optical rotation: (1 g/dL, MeOH)=−40

MS: 370.3 ES+ (C₁₉H₃₅NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.00 (d, J=7.93 Hz, 6H); 0.82 (s, 9H);1.48 (s, 9H); 1.79-1.87 (m, 3H); 1.98 (s, 3H); 3.79-4.06 (m, 4H);4.36-4.58 (m, 1H).

Intermediate 191: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3,4-dimethyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 190, 2.39 g, 6.47 mmol) following the procedure describedfor Intermediate 8. Silica gel chromatography (0%-20% ethylacetate/hexanes) afforded the desired product as a colorless oily solid(1.73 g, 72%).

MS: 372.3 ES+ (C₁₉H₃₇NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.01-0.12 (m, 6H); 0.83-0.94 (m, 9H);1.48 (s, 9H); 1.69 (s, 3H); 1.84 (s, 3H); 3.34 (d, J=12.84 Hz, 1H); 3.68(d, J=8.12 Hz, 2H); 3.87 (br. s., 1H); 4.00-4.29 (m, 2H).

Intermediate 192: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 191, 1.73 g, 4.66 mmol) andN-(allyloxy)-2-nitrobenzenesulfonamide (Intermediate 9, 1.26 g, 4.89mmol) following the procedure described for Intermediate 10. Silica gelchromatography (0%-20% ethyl acetate/hexanes) afforded the desiredproduct as an off-white solid (3.15 g, 100%).

MS: 612.3 ES+ (C₂₈H₄₅N₃O₈SSi)

¹H NMR (300 MHz, CHLOROFORM-d) δ: −0.05-0.08 (m, 6H); 0.78-0.91 (m, 9H);1.31-1.89 (m, 15H); 3.17-3.86 (m, 4H); 4.17-4.31 (m, 1H); 4.39 (br. s.,1H); 4.57 (dt, J=6.23, 1.23 Hz, 2H); 5.05-5.43 (m, 2H); 5.94 (ddt,J=17.09, 10.48, 6.33, 6.33 Hz, 1H); 7.50-7.64 (m, 1H); 7.66-8.00 (m,1H); 8.02-8.32 (m, 2H).

Intermediate 193: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 192, 3.15 g, 5.15 mmol) following the procedure describedfor Intermediate 18. Two silica gel chromatography (0%-100% ethylacetate/hexanes) afforded the desired product as a white foamy solid(1.68 g, 65.6%).

MS: 498.2 ES+ (C₂₂H₃₁N₃O₈S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.15-1.92 (m, 15H); 2.98-3.46 (m, 1H);3.55-3.96 (m, 3H); 4.01-4.35 (m, 3H); 4.57 (d, J=6.80 Hz, 1H); 5.09-5.35(m, 2H); 5.54-5.91 (m, 1H); 7.61 (d, J=7.37 Hz, 1H); 7.67-7.85 (m, 2H);8.08-8.22 (m, 1H).

Intermediate 194:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3,4-dimethyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 193, 1.68 g, 3.38 mmol) following the procedure describedfor Intermediate 19. The workup yielded organics that were dried overmagnesium sulfate, filtered and concentrated crude material to afford alight orange foam (1.48 g, 86%).

MS: 512.2 ES+ (C₂₂H₂₉N₃O₉S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.22-1.61 (m, 12H); 1.86 (s, 3H);3.15-3.42 (m, 1H); 3.82-4.07 (m, 1H); 4.09-4.34 (m, 2H); 4.92 (s, 1H);5.09-5.24 (m, 2H); 5.31 (s, 1H); 5.56-5.84 (m, 1H); 7.52-7.67 (m, 1H);7.69-7.91 (m, 2H); 8.13 (dd, J=7.84, 1.42 Hz, 1H).

Intermediate 195: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3,4-dimethyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 194, 1.48 g, 2.89 mmol) following the proceduredescribed for Intermediate 20. Silica gel chromatography (0%-80% ethylacetate/hexanes) afforded the desired product as a light yellow foamysolid (1.41 g, 95%).

Optical rotation: (0.4 g/dL, MeOH)=−51

MS: 511.2 ES+ (C₂₂H₃₀N₄O₈S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.15-1.48 (m, 9H); 1.51-1.85 (m, 6H);2.93-3.36 (m, 2H); 4.02-4.34 (m, 2H); 4.83 (br. s., 1H); 5.04-5.40 (m,3H); 5.51-5.77 (m, 1H); 7.60 (d, J=7.36 Hz, 1H); 7.65-7.85 (m, 2H);7.99-8.18 (m, 1H).

Intermediate 196:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3,4-dimethyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3,4-dimethyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 195, 1.36 g, 2.66 mmol) following the procedure describedfor Intermediate 21. The workup yielded an organic layer that was driedover magnesium sulfate, filtered and concentrated to afford the desiredproduct as a light pink solid (0.82 g, 75%).

MS: 411.1 ES+ (C₁₇H₂₂N₄O₆S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.87 (s, 3H); 1.98 (s, 3H); 2.67-2.89(m, 2H); 3.66 (s, 1H); 3.92-4.31 (m, 2H); 4.51 (dd, J=11.33, 6.04 Hz,1H); 5.15-5.34 (m, 2H); 5.67-5.93 (m, 1H); 7.61 (dd, J=7.84, 1.42 Hz,1H); 7.68-7.94 (m, 2H); 8.05-8.18 (m, 1H).

Intermediate 197:(2S,5R)-5-(allyloxyamino)-3,4-dimethyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3,4-dimethyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 196, 0.82 g, 2.00 mmol) following the procedure describedfor Intermediate 12. Silica gel chromatography (0%-10%methanol/dichloromethane) afforded the desired product as a white solid(0.31 g, 68.9%).

MS: 226.2 ES+ (C₁₁H₁₉N₃O₂)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.80 (s, 3H); 1.86 (s, 3H); 2.83-2.97(m, 1H); 3.12 (br. s., 1H); 3.22 (d, J=13.79 Hz, 1H); 3.76 (br. s., 1H);4.20 (dt, J=6.00, 1.25 Hz, 2H); 5.16-5.35 (m, 2H); 5.42 (br. s., 1H);5.95 (ddt, J=17.28, 10.34, 6.02, 6.02 Hz, 1H); 7.06-7.20 (m, 1H).

Intermediate 198:(2S,5R)-6-(allyloxy)-3,4-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

The title compound was prepared from(2S,5R)-5-(allyloxyamino)-3,4-dimethyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 197, 0.31 g, 1.38 mmol) following the procedure describedfor Intermediate 13. Silica gel chromatography (0%-70% ethylacetate/hexanes) afforded the desired product as a white solid (0.23 g,66.5%).

MS: 252.2 ES+ (C₁₂H₁₇N₃O₃)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.76-1.82 (m, 3H); 1.83-1.90 (m, 3H);3.10-3.33 (m, 1H); 3.66 (d, J=2.64 Hz, 1H); 4.23 (s, 1H); 4.42 (qdt,J=12.20, 12.20, 12.20, 6.35, 1.09, 1.09 Hz, 2H); 5.24-5.51 (m, 3H); 6.02(ddt, J=17.00, 10.53, 6.35, 6.35 Hz, 1H); 6.61 (br. s., 2H).

Intermediate 199: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3,4-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-3,4-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 198, 0.227 g, 0.9 mmol) following the procedure describedfor Intermediate 17. Silica gel chromatography (0%-100%acetone/dichloromethane) afforded the desired product as a light yellowoily solid (0.5 g, 93%).

MS: 290 ES−, 303 ES+ (C₉H₁₂N₃O₆S, C₂₁H₂₀P)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.77 (s, 3H); 1.86-1.95 (m, 3H);2.28-2.39 (m, 3H); 3.14 (d, J=10.76 Hz, 1H); 3.46 (dd, J=10.76, 3.02 Hz,1H); 4.29 (d, J=2.83 Hz, 1H); 5.38 (br. s., 1H); 6.51-6.80 (m, 1H);7.20-7.54 (m, 6H); 7.58-7.91 (m, 10H).

Example 22(2S,5R)-2-carbamoyl-3-ethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-en-1-yl)phosphonium(2S,5R)-2-carbamoyl-3-ethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 216, 0.77 g, 1.30 mmol) according to the procedurefor Example 1. Fractions 5-7 were combined and lyophilized separatelyfrom fractions 8-17 which were yellow. The title compound was obtainedfrom fractions 5-7 (0.275 g, 67.5%) as an off-white solid.

MS: 292 ES+ (C₉H₁₃N₃O₆S)

¹H NMR (300 MHz, DMSO-d6) δ: 0.94 (t, J=7.54 Hz, 3H) 1.59-2.19 (m, 2H)3.07 (dd, J=10.93, 1.88 Hz, 1H) 3.73 (d, J=10.55 Hz, 1H) 4.06 (dd,J=4.52, 2.26 Hz, 1H) 4.15 (s, 1H) 6.03 (d, J=4.52 Hz, 1H) 7.29 (br. s.,1H) 7.83 (br. s., 1H).

The Intermediates for Example 22 were Prepared as Follows Intermediate200: (R)-tert-butyl4-(3-hydroxypentan-3-yl)-2,2-dimethyloxazolidine-3-carboxylate

A suspension of cerium (III) chloride (119 g, 482.07 mmol) in THF (350mL) at room temperature was stirred vigorously for 2 hours. Thesuspension was cooled to −78° C. and ethylmagnesium bromide (482 mL,482.07 mmol) was added dropwise. The mixture was stirred at −78° C. for1.5 hours. (R)-3-tert-butyl 4-methyl2,2-dimethyloxazolidine-3,4-dicarboxylate (Aldrich, 25 g, 96.41 mmol) inTHF (100 mL) was then added dropwise at −78° C. The reaction was stirredat −78° C. for 30 minutes and then warmed to 0° C. for 15 minutes. Thereaction was quenched with saturated NH₄Cl, diluted further with waterand extracted twice with ether. The organics were dried over magnesiumsulfate, filtered and concentrated. Silica gel chromatography (0%-20%ethyl acetate/hexanes) afforded the title compound (26.4 g, 95%) as acolorless oil.

MS: 288 ES+ (C₁₅H₂₉NO₄)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.77-0.98 (m, 6H) 1.24-1.81 (m, 19H)3.86 (d, J=8.29 Hz, 1H) 3.95-4.07 (m, 1H) 4.17 (d, J=7.35 Hz, 1H) 4.97(br. s., 1H).

Intermediate 201: (S)-tert-butyl2,2-dimethyl-4-(pent-2-en-3-yl)oxazolidine-3-carboxylate

To a solution of (R)-tert-butyl4-(3-hydroxypentan-3-yl)-2,2-dimethyloxazolidine-3-carboxylate(Intermediate 200, 26.4 g, 91.86 mmol) in dichloromethane (350 mL) at 0°C. was added triethylamine (128 mL, 918.60 mmol) and then slowly addedmethanesulfonyl chloride (35.8 mL, 459.30 mmol). The yellow reactionmixture was warmed to room temperature and stirred overnight for ˜16 h.The reaction mixture was diluted with diethyl ether and poured intowater. The organic layer was washed with 10% citric acid, saturatedsodium bicarbonate, saturated sodium chloride, dried over sodiumsulfate, filtered, and concentrated under reduced pressure to give abrown oil. Purification by flash column chromatography (0-20% ethylacetate/hexanes) afforded the title compound (17.70 g, 71.5%) as verypale yellow oil.

MS: 270 ES+ (C₁₅H₂₇NO₃)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.02 (q, J=7.47 Hz, 3H) 1.38-1.54 (m,12H) 1.61-1.73 (m, 6H) 1.83-2.29 (m, 2H) 3.59-3.75 (m, 1H) 4.03-4.16 (m,1H) 4.18-5.01 (m, 1H) 5.32 (br. s., 1H).

Intermediate 202: (S)-tert-butyl3-ethyl-1-hydroxypent-3-en-2-ylcarbamate

A solution of (S)-tert-butyl2,2-dimethyl-4-(pent-2-en-3-yl)oxazolidine-3-carboxylate (Intermediate201, 15.815 g, 58.71 mmol) and p-toluenesulfonic acid monohydrate (3.35g, 17.61 mmol) in methanol (100 mL) was heated to 85° C. for 3 h. Thereaction mixture was cooled to room temperature and triethylamine (8.18mL, 58.71 mmol) and di-tert-butyl dicarbonate (6.82 mL, 29.35 mmol) wereadded. The reaction mixture was stirred at room temperature overnight.The reaction mixture was concentrated under reduced pressure to give ayellow oil. Purification by flash column chromatography (0-100% ethylacetate/hexanes) afforded the title compound (10.12 g, 75%) as a whitesolid.

MS: 230 ES+ (C₁₂H₂₃NO₃)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.99-1.08 (m, 3H) 1.46 (q, J=2.07 Hz,9H) 1.64-1.75 (m, 3H) 1.92-2.22 (m, 3H) 3.54-3.76 (m, 2H) 4.15-4.89 (m,2H) 5.43 (q, J=6.84 Hz, 1H).

Intermediate 203: (S,E)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-ylcarbamate

To a solution of (S)-tert-butyl 3-ethyl-1-hydroxypent-3-en-2-ylcarbamate(Intermediate 202, 11.855 g, 51.70 mmol) in DCM (120 mL) was addedimidazole (5.28 g, 77.55 mmol), DMAP (1.263 g, 10.34 mmol) and TBDMS-Cl(9.35 g, 62.04 mmol). The reaction mixture was stirred at roomtemperature overnight. The reaction mixture was diluted with ethyl etherand washed with brine. The organic layer was dried over sodium sulfate,filtered, and concentrated under reduced pressure. Purification by flashcolumn chromatography (0-50% ethyl acetate/hexanes) afforded the titlecompound (17.20 g, 97%) as a clear oil.

MS: 344 ES+ (C₁₈H₃₇NO₃Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.06 (d, J=1.13 Hz, 6H) 0.89 (dd,J=2.83, 1.32 Hz, 9H) 0.96-1.08 (m, 3H) 1.45 (s, 9H) 1.54-1.76 (m, 3H)1.89-2.20 (m, 2H) 3.50-3.76 (m, 2H) 3.93-4.71 (m, 1H) 4.87 (br. s., 1H)5.27-5.46 (m, 1H).

Intermediate 204:(S)-1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-amine

To a solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-ylcarbamate(Intermediate 203, 17.2 g, 50.06 mmol) in DCM (300 mL) at roomtemperature was added zinc bromide (45.1 g, 200.25 mmol). The reactionmixture was stirred for 2 days. The reaction mixture was filteredthrough fitted funnel and washed with saturated sodium bicarbonate. Theaqueous layer was extracted two more times with DCM and the organicswere combined, dried over magnesium sulfate, filtered and concentratedto afford the title compound (12.19 g, 100%) as a yellow oil.

MS: 244 ES+ (C₁₃H₂₉NOSi)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.02-0.12 (m, 6H) 0.86-0.94 (m, 9H)0.95-1.05 (m, 3H) 1.60-1.72 (m, 3H) 1.87-2.22 (m, 2H) 3.29-3.57 (m, 2H)3.58-4.00 (m, 1H) 5.24-5.51 (m, 1H).

Intermediate 205: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate

A mixture of (S)-1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-amine(Intermediate 204, 14.14 g, 58.08 mmol) and cesium carbonate (18.92 g,58.08 mmol) in DMF (140 mL) was stirred at room temperature for 1 hour.2-bromo-N-methoxy-N-methylacetamide (Intermediate 4, 9.61 g, 52.80 mmol)was added and the reaction mixture was stirred at room temperature for 6hours. Di-tert-butyl dicarbonate (12.74 mL, 55.44 mmol) was added andthe reaction mixture was stirred at room temperature overnight.

The reaction mixture was diluted with ethyl acetate and filtered toremove the inorganic salts. The filtrate was concentrated and theresidue was taken up in ether and washed with aqueous saturated sodiumbicarbonate, water, aqueous 5% citric acid, water and brine. Theorganics were dried over magnesium sulfate, filtered and concentrated.Silica gel chromatography (0%-30% ethyl acetate/hexanes) afforded thetitle compound (13.71 g, 58.4%) as a light yellow oil.

MS: 445 ES+ (C₂₂H₄₄N₂O₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: −0.05-0.05 (m, 6H) 0.83 (s, 9H) 0.87-0.98(m, 3H) 1.27-1.43 (m, 9H) 1.59 (dd, J=8.57, 7.06 Hz, 3H) 1.84-2.15 (m,2H) 3.01-3.11 (m, 3H) 3.63 (d, J=6.03 Hz, 3H) 3.69-3.92 (m, 3H)3.98-4.22 (m, 1H) 4.43-4.91 (m, 1H) 5.27-5.42 (m, 1H).

Intermediate 206: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-yl(2-oxopent-3-enyl)carbamate

The title compound was prepared from (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 205, 13.7 g, 30.81 mmol) according to the proceduredescribed for Intermediate 236. The reaction was quenched with saturatedNH₄Cl, diluted further with water and extracted twice with ether. Theorganics were dried over magnesium sulfate, filtered and concentrated.Silica gel chromatography (0%-45% ethyl acetate/hexanes) afforded thetitle compound (10.95 g, 83%) as a colorless oil.

MS: 426 ES+ (C₂₃H₄₃NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: −0.02-0.07 (m, 6H) 0.83-0.91 (m, 9H)1.00 (t, J=7.35 Hz, 3H) 1.33-1.51 (m, 9H) 1.57-1.74 (m, 3H) 1.82-1.92(m, 3H) 1.92-2.21 (m, 2H) 3.65-3.97 (m, 3H) 3.97-4.34 (m, 1H) 4.54-5.08(m, 1H) 5.25-5.44 (m, 1H) 6.12-6.35 (m, 1H) 6.79-7.00 (m, 1H).

Intermediate 207: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-ethyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-ethylpent-3-en-2-yl(2-oxopent-3-enyl)carbamate(Intermediate 206, 10.95 g, 25.72 mmol) according to the proceduredescribed for Intermediate 7, except the reaction was heated at 86° C.for 6 days. Silica gel chromatography (0-10% ethyl acetate/hexanes)afforded the title compound (5.15 g, 54.1%) as a light yellow oil.

MS: 370 ES+ (C₁₉H₃₅NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: −0.18-0.09 (m, 6H) 0.70-0.94 (m, 9H)1.17 (t, J=7.35 Hz, 3H) 1.48 (s, 9H) 2.19-2.43 (m, 2H) 3.68-4.06 (m, 3H)4.28-4.78 (m, 2H) 6.08 (s, 1H).

Intermediate 208: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-ethyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-ethyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 207, 5.146 g, 13.92 mmol) according to the proceduredescribed for Intermediate 8, except the mixture was stirred at ambienttemperature for 1 h. The mixture was concentrated and diluted withaqueous NH₄Cl, water and EtOAc. The organic layer was separated andwashed three times with brine, dried over Na₂SO₄, filtered andconcentrated to afford the title compound (5.19 g, 100%) as a cloudyoil.

MS: 372 ES+ (C₁₉H₃₇NO₄Si)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.06 (s, 6H) 0.80-0.93 (m, 9H)1.05-1.14 (m, 3H) 1.48 (s, 9H) 1.98-2.17 (m, 2H) 2.83-3.53 (m, 2H) 3.74(d, J=11.30 Hz, 2H) 4.06-4.35 (m, 3H) 5.82 (br. s., 1H).

Intermediate 209: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-ethyl-5,6-dihydropyridine-1(2H)-carboxylate

To a stirred suspension of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-ethyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 208, 6.36 g, 17.12 mmol),N-(allyloxy)-2-nitrobenzenesulfonamide (Intermediate 9) (8.84 g, 34.23mmol) and triphenylphosphine (13.47 g, 51.35 mmol) in toluene (25 mL) at0° C. was added dropwise (E)-diisopropyl diazene-1,2-dicarboxylate (9.95ml, 51.35 mmol). The reaction mixture was warmed to room temperature andstirred for 30 minutes. Silica gel chromatography (0-40% ethylacetate/hexanes) afforded the title compound (7.00 g, 66.8%) as a yellowoil.

MS: 612 ES+ (C₂₈H₄₅N₃O₈SSi)

Intermediate 210: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-ethyl-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-ethyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 209, 7 g, 11.44 mmol) following the procedure describedfor Intermediate 18. Silica gel chromatography (30%-90% ethylacetate/hexanes) afforded the title compound (4.93 g, 87%) as an tanfoam.

MS: 498 ES+ (C₂₂H₃₁N₃O₈S)

Intermediate 211:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-ethyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-ethyl-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 210, 2.35 g, 4.72 mmol) following the procedure describedfor Intermediate 19. The title compound was obtained as a tan foam (1.89g, 78%).

MS: 512 ES+ (C₂₂H₂₉N₃O₉S)

Intermediate 212: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-ethyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-ethyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 211, 1.89 g, 3.69 mmol) following the proceduredescribed for Intermediate 20. Silica gel chromatography (0%-80% ethylacetate/hexanes) afforded the title compound (1.445 g, 77%) as a beigefoam.

MS: 511 ES+ (C₂₂H₂₉N₃O₉S)

Intermediate 213:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-ethyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-ethyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 212, 1.44 g, 2.82 mmol) following the procedure describedfor Intermediate 21, except the reaction mixture was stirred over theweekend at room temperature. The reaction mixture was diluted withdichloromethane and washed with saturated sodium bicarbonate. Anemulsion formed and the aqueous was extracted three to four more timeswith DCM. The organics were washed with brine, dried over magnesiumsulfate, filtered and concentrated to afford the title compound (0.870g, 75%) as a yellow foam.

MS: 411 ES+ (C₁₇H₂₂N₄O₆S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.04 (t, J=7.54 Hz, 3H) 2.17-2.35 (m,1H) 2.35-2.52 (m, 1H) 2.94 (d, J=5.27 Hz, 2H) 3.79 (s, 1H) 4.29 (d,J=3.01 Hz, 1H) 4.35-4.56 (m, 2H) 5.18-5.39 (m, 3H) 5.55 (br. s., 1H)5.74-5.94 (m, 1H) 7.04 (br. s., 1H) 7.55-7.67 (m, 1H) 7.69-7.86 (m, 2H)8.06-8.23 (m, 1H).

Intermediate 214:(R)-5-(allyloxyamino)-3-ethyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-ethyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 213, 0.87 g, 2.12 mmol) following the procedure describedfor Intermediate 12. Silica gel chromatography (0%-8%methanol/dichloromethane) afforded the title compound (0.355 g, 74.3%)as an off-white solid upon drying on high-vacuum.

MS: 226 ES+ (C₁₁H₁₉N₃O₂)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.07 (t, J=7.54 Hz, 3H) 2.10-2.48 (m,3H) 2.90-3.08 (m, 2H) 3.43 (d, J=2.26 Hz, 1H) 3.84 (s, 1H) 4.21 (dd,J=6.78, 1.51 Hz, 2H) 5.16-5.67 (m, 5H) 5.95 (ddt, J=17.14, 10.74, 6.03,6.03 Hz, 1H) 7.08 (br. s., 1H).

Intermediate 215:(2S,5R)-6-(allyloxy)-3-ethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of(R)-5-(allyloxyamino)-3-ethyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 214, 0.35 g, 1.55 mmol) and diisopropylethyl amine (1.082mL, 6.21 mmol) in acetonitrile (110 mL) at 0° C. was added triphosgene(0.157 g, 0.53 mmol) as a solution in acetonitrile (4 mL). Thetriphosgene solution was added at a rate of 4 mL/hour. Once addition wascomplete the reaction was stirred at 0° C. for ˜1 hour. The reactionmixture was diluted with ethyl acetate, then washed with saturatedsodium bicarbonate and brine. The aqueous washes were extracted oncewith dichloromethane. The combined organic extracts were dried overmagnesium sulfate, filtered and concentrated. Silica gel chromatography(0%-100% ethyl acetate/hexanes) afforded the title compound (0.340 g,87%) as a colorless oil.

MS: 252 ES+ (C₁₂H₁₇N₃O₃)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 1.05 (t, J=7.16 Hz, 3H) 2.08-2.47 (m,2H) 3.15-3.34 (m, 2H) 3.85 (dd, J=5.27, 3.01 Hz, 1H) 4.31-4.50 (m, 3H)5.27-5.51 (m, 3H) 5.93-6.15 (m, 2H) 6.65 (br. s., 1H).

Intermediate 216: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-ethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-3-ethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 215, 340 mg, 1.35 mmol) following the procedure describedfor Intermediate 17. Silica gel chromatography (0%-100% acetone/DCM)afforded the title compound (770 mg, 96%) as a yellow foam.

MS: 303 ES+ (C₂₁H₂₀P) and 290 ES− (C₉H₁₂N₃O₆S)

¹H NMR (300 MHz, CHLOROFORM-d) δ: 0.99 (t, J=7.16 Hz, 3H) 2.05-2.16 (m,1H) 2.07-2.16 (m, 1H) 2.34 (td, J=4.33, 1.88 Hz, 3H) 2.36-2.47 (m, 1H)3.13 (d, J=10.55 Hz, 1H) 3.45-3.55 (m, 1H) 4.31 (s, 1H) 4.42 (dd,J=5.27, 3.01 Hz, 1H) 5.40 (br. s., 1H) 6.18 (d, J=5.27 Hz, 1H) 6.53-6.81(m, 2H) 7.35-7.47 (m, 2H) 7.62-7.87 (m, 13H).

Example 23(2S,5R)-4-(2-aminoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate

To a suspension of(2S,5R)-4-(2-(tert-butoxycarbonylamino)ethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate (Intermediate 229, 0.060 g, 0.15 mmol) indichloromethane (2 mL) at 0° C. was added trifluoroacetic acid (0.100mL). After 15 minutes the solvent was removed in vacuo and the residuewas dried under high vacuum. Purified by reverse phase chromatographyusing acetonitrile/water to afford the title compound as a white solidafter lyophilization (0.006 g, 13%).

MS: 305 ES− (C₉H₁₄N₄O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.19-2.33 (m, 2H) 2.71-2.87 (m, 1H) 2.93(d, J=9.80 Hz, 1H) 3.13 (br. s., 2H) 3.89-4.14 (m, 2H) 5.49 (br. s., 1H)7.27 (br. s., 1H) 7.52 (d, J=11.30 Hz, 3H).

The Intermediates for Example 23 were Prepared as Follows Intermediate217: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared as a colorless oil (13.7 g, 88%),according to the procedure described for Intermediate 79, from(S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-(hydroxymethyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 117).

MS: 374 ES+ (C₁₈H₃₅NO₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.00 (s, 6H) 0.75-0.89 (m, 9H) 1.37 (s, 9H)3.59 (d, J=5.27 Hz, 2H) 3.88-4.00 (m, 3H) 4.15-4.36 (m, 1H) 4.57 (t,J=5.65 Hz, 1H) 5.00 (d, J=5.27 Hz, 1H) 5.62 (d, J=2.26 Hz, 1H).

Intermediate 218: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-((ethoxycarbonyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 217, 13.7 g, 36.67 mmol) in dichloromethane (20 mL) at 0°C. was added pyridine (5.93 mL, 73.35 mmol) followed by dropwiseaddition of ethyl chloroformate (3.87 mL, 40.34 mmol). The reaction wasallowed to warm to room temperature over 3 hours. Diluted withdichloromethane and washed with saturated NH₄Cl. Extracted withdichloromethane three times, combined and dried over Na₂SO₄. Removedsolvent to give title compound as a yellow oil (14.5 g, 89%).

MS: 446 ES+ (C₂₁H₃₉NO₇Si)

Intermediate 219: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-((ethoxycarbonyloxy)methyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 218, 14.5 g, 32.54 mmol) was taken up in nitromethane (100ml, 1854.52 mmol) and degassed for 5 minutes. Pd₂(dba)₃ (1.49 g, 1.63mmol) was added and the reaction mixture was stirred under nitrogen atroom temperature for 18 hrs. Reaction mixture was dried directly ontosilica and purified by flash chromatography using 0-50% EtOAc/Hexanes.The title compound was obtained as a yellow oil (6 g, 44.3%).

MS: 417 ES+ (C₁₉H₃₆N₂O₆Si)

Intermediate 220: (2S,5R)-tert-butyl5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared as a pale yellow oil (5 g, 30%)according to the procedure described for Intermediate 80, from(2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-5-hydroxy-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate (Intermediate 219, 6.73 g, 16.3 mmol) andN-(benzyloxy)-2-nitrobenzenesulfonamide (7.40 g, 24.00 mmol).

MS: 707 ES+ (C₃₂H₄₆N₄O₁₀SSi)

Intermediate 221: (2S,5R)-tert-butyl5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate

(2S,5R)-tert-butyl5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 220, 5 g, 7.07 mmol) was taken up in methanol (150 mL)cooled to 0° C. and acetyl chloride (0.075 mL, 1.06 mmol) added. Thereaction mixture was stirred for 3 hours at 0° C. The solvent wasremoved in vacuo and the residue was taken up in EtOAc, washed withsaturated sodium bicarbonate, dried over Na₂SO₄, filtered andconcentrated to afford the title compound as an oil (4.1 g, 98%).

MS: 593 ES+ (C₂₆H₃₂N₄O₁₀S)

Intermediate 222:(2S,5R)-5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-(2-nitroethyl)-1,2,5,6-tetrahydropyridine-2-carboxylicacid

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-2-(hydroxymethyl)-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 221, 4.1 g, 6.92 mmol) according to the proceduredescribed for Intermediate 19. The desired product was obtained as anoff-white foam (4.0 g, 98%).

MS: 606 ES+ (C₂₆H₃₀N₄O₁₁S)

Intermediate 223: (2S,5R)-tert-butyl5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from(2S,5R)-5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-(2-nitroethyl)-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 222, 4.0 g, 6.59 mmol) according to the proceduredescribed for Intermediate 20. The desired product was obtained as anoff-white foam (2 g, 50%).

MS: 606 ES+ (C₂₆H₃₁N₅O₁₀S)

Intermediate 224: (2S,5R)-tert-butyl5-(benzyloxyamino)-2-carbamoyl-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The desired product was prepared from (2S,5R)-tert-butyl5-(N-(benzyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 223, 2 g, 3.30 mmol) following the procedure described forIntermediate 22. The desired product was obtained as yellow oil (0.8 g,68%).

MS: 421 ES+ (C₂₀H₂₈N₄O₆)

Intermediate 225:(2S,5R)-5-(benzyloxyamino)-4-(2-nitroethyl)-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from (2S,5R)-tert-butyl5-(benzyloxyamino)-2-carbamoyl-4-(2-nitroethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 224, 0.800 g, 3.30 mmol) according to the proceduredescribed for Intermediate 21. The desired product was obtained as ayellow oil (0.6 g, 98%).

MS: 321 ES+ (C₁₅H₂₀N₄O₄)

Intermediate 226:(2S,5R)-6-(benzyloxy)-4-(2-nitroethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

The title compound was prepared from(2S,5R)-5-(benzyloxyamino)-4-(2-nitroethyl)-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 225, 0.600 g, 1.87 mmol) according to the proceduredescribed for Intermediate 16. The desired product was obtained as alight yellow oil (0.510 g, 79%).

MS: 347 ES+ (C₁₆H₁₈N₄O₅)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.69 (t, J=6.78 Hz, 2H) 3.17 (s, 2H) 3.88(s, 1H) 4.09-4.18 (m, 1H) 4.55-4.70 (m, 2H) 4.91 (d, J=3.01 Hz, 2H) 5.56(br. s., 1H) 7.30 (br. s., 1H) 7.35-7.46 (m, 5H) 7.50 (br. s., 1H).

Intermediate 227: tert-butyl2-((2S,5R)-6-(benzyloxy)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-4-yl)ethylcarbamate

(2S,5R)-6-(benzyloxy)-4-(2-nitroethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 226, 0.420 g, 1.21 mmol) was taken up in ethanol(absolute, 99.5%) (20 mL), and zinc dust (1.983 g, 30.32 mmol) was addedfollowed by acetic acid (2.78 mL, 48.51 mmol). The reaction mixture wasstirred at ambient temperature for 1 hour. DIPEA (2.118 mL, 12.13 mmol)was added followed by BOC-anhydride (0.845 mL, 3.64 mmol). The reactionmixture was stirred for 2 hours, then diluted with DCM, washed withsaturated bicarbonate and concentrated to dryness. Flash chromatography,0-10% MeOH/DCM, afforded the title compound as a clear oil (0.31 g,61%).

MS: 417 ES+ (C₂₁H₂₈N₄O₅)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.10-1.26 (m, 3H) 1.28-1.46 (m, 9H) 2.17(d, J=6.03 Hz, 1H) 2.79-3.16 (m, 2H) 3.18-3.26 (m, 1H) 3.55 (br. s., 1H)4.21 (br. s., 1H) 4.71 (s, 3H) 4.76-4.95 (m, 2H) 5.53 (d, J=2.26 Hz, 1H)7.23-7.46 (m, 4H).

Intermediate 228: tert-butyl2-((2S,5R)-2-carbamoyl-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-4-yl)ethylcarbamate

To a solution of tert-butyl2-((2S,5R)-6-(benzyloxy)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-4-yl)ethylcarbamate(Intermediate 227, 0.300 g, 0.72 mmol) in EtOAc (5 mL) was added 10%Pd/C (0.077 g, 0.72 mmol). The reaction mixture was stirred underhydrogen for 30 minutes. The reaction mixture was filtered throughcelite and washed with EtOAc. The solvent was removed in vacuo to givethe title compound as a white solid (0.23 g, 98%).

MS: 327 ES+ (C₁₄H₂₂N₄O₅)

Intermediate 229:(2S,5R)-4-(2-(tert-butoxycarbonylamino)ethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate

To a solution of tert-butyl2-((2S,5R)-2-carbamoyl-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-4-yl)ethylcarbamate(Intermediate 228, 0.230 g, 0.70 mmol) in pyridine (5 mL) was addedpyridine/sulfur trioxide complex (0.673 g, 4.23 mmol). The reaction wasstirred overnight at room temperature. The solvent was removed in vacuo,and the residue was purified by reverse phase chromatography usingacetonitrile/water. Desired fractions were combined and lyophilized toafford the title compound as a while solid (0.23 g, 80%) as a whitesolid.

MS: 405 ES− (C₁₄H₂₂N₄O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.36 (s, 9H) 2.17 (t, J=6.40 Hz, 2H)2.90-3.27 (m, 4H) 4.08 (d, J=18.84 Hz, 2H) 5.43 (d, J=2.26 Hz, 1H) 6.61(br. s., 1H) 7.25 (br. s., 1H) 7.47 (br. s., 1H) 7.80-7.95 (m, 1H) 8.38(t, J=7.54 Hz, 1H) 8.83 (d, J=5.27 Hz, 1H).

Example 24(2S,5R)-2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 246, 0.39 g, 0.64 mmol) following the proceduredescribed for Example 1. The desired product was obtained as anoff-white solid (158 mg, 75%).

Optical rotation: (0.1 g/dL, MeOH)=−50

MS: 302 ES− (C₁₀H₁₃N₃O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.38 (m, 2H); 0.59 (m, 2H); 1.19 (m, 1H);3.03 (m, 1H); 3.78 (m, 1H); 4.02 (m, 1H); 4.20 (m, 1H); 5.91 (m, 1H);7.28 (bs, 1H); 7.85 (bs, 1H).

The Intermediates for Example 24 were Prepared as Follows Intermediate230: (R)-tert-butyl4-(cyclopropyl(hydroxy)methyl)-2,2-dimethyloxazolidine-3-carboxylate

To a solution of (R)-tert-butyl4-formyl-2,2-dimethyloxazolidine-3-carboxylate (Aldrich, 12.44 g, 54.26mmol) in THF (150 mL) at −78° C. was added cyclopropylmagnesium bromide(217 mL, 108.52 mmol) dropwise. The reaction mixture was allowed to warmto room temperature and stir overnight. The reaction was quenched withwater and diluted with ethyl acetate and brine. The resulting emulsionwas filtered through celite and the layers separated. The organics weredried over magnesium sulfate, filtered and concentrated. Silica gelchromatography (0%-20% ethyl acetate/hexanes) afforded the titlecompound as a light yellow oil (12.47 g, 85%).

¹H NMR (300 MHz, DMSO-d₆) δ: 0.16 (m, 2H); 0.37 (m, 2H); 0.82 (m, 1H);1.45 (m, 15H); 2.87 (m, 1H); 3.86 (m, 2H); 3.97 (m, 1H); 4.74 (m, 1H).

Intermediate 231: (R)-tert-butyl4-(cyclopropanecarbonyl)-2,2-dimethyloxazolidine-3-carboxylate

To a solution of (R)-tert-butyl4-(cyclopropyl(hydroxy)methyl)-2,2-dimethyloxazolidine-3-carboxylate(Intermediate 230, 12.47 g, 45.95 mmol) in DCM (300 mL) at roomtemperature was added Dess-Martin periodinane (29.2 g, 68.93 mmol). Thereaction mixture was stirred overnight then diluted with ethyl acetateand washed with saturated sodium bicarbonate. An emulsion formed and wasfiltered through celite. The layers were separated and the organicswashed with brine. The organics were dried over magnesium sulfate,filtered and concentrated. Silica gel chromatography (0%-20% ethylacetate/hexanes) afforded the title compound as a colorless oil (11.15g, 90%).

¹H NMR (300 MHz, DMSO-d₆) δ: 0.90 (m, 4H); 1.38 (m, 12H); 1.54 (m, 3H);2.12 (m, 1H); 3.94 (m, 1H); 4.18 (m, 1H); 4.56 (m, 1H).

Intermediate 232: (S)-tert-butyl4-(1-cyclopropylvinyl)-2,2-dimethyloxazolidine-3-carboxylate

To a suspension of potassium tert-butoxide (9.29 g, 82.80 mmol) in ether(250 mL) at room temperature was added methyltriphenylphosphoniumbromide (29.6 g, 82.80 mmol). The mixture turned bright yellow and washeated to 40° C. for 1 hour. The mixture was cooled to room temperatureand a solution of (R)-tert-butyl4-(cyclopropanecarbonyl)-2,2-dimethyloxazolidine-3-carboxylate(Intermediate 231, 11.15 g, 41.40 mmol) in ether (30 mL) was added andthe reaction mixture was stirred for 2 hours. The reaction was quenchedwith water (10 mL) and the layers were separated. The aqueous wasextracted once with ether. The combined organic extracts were dried overmagnesium sulfate, filtered and concentrated. Silica gel chromatography(0%-15% ethyl acetate/hexanes) afforded the title compound as acolorless oil (9.84 g, 89%).

¹H NMR (300 MHz, DMSO-d₆) δ: 0.42 (m, 2H); 0.65 (m, 2H); 1.43 (m, 16H);3.76 (m, 1H); 4.09 (m, 1H); 4.27 (m, 1H); 4.66 (m, 2H).

Intermediate 233: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-ylcarbamate

To a solution of (S)-tert-butyl4-(1-cyclopropylvinyl)-2,2-dimethyloxazolidine-3-carboxylate(Intermediate 232, 8.25 g, 30.86 mmol) in methanol (100 mL) at roomtemperature was added p-toluenesulfonic acid monohydrate (1.174 g, 6.17mmol). The reaction mixture was heated to 80° C. overnight. Another 0.2eq of p-toluenesulfonic acid monohydrate was added. Continue heating at80° C. for 2 hours. The reaction mixture was cooled to room temperature.Triethylamine (4.29 mL, 30.86 mmol) and di-tert-butyl dicarbonate (3.37g, 15.43 mmol) were added. The reaction mixture was stirred over theweekend then concentrated. The residue was dissolved in ethyl acetateand washed once with saturated sodium bicarbonate. The combined organicextracts were dried over magnesium sulfate, filtered and concentrated.The resulting oil was dissolved in DCM (100 mL). Imidazole (2.73 g,40.11 mmol), 4-dimethylaminopyridine (0.754 g, 6.17 mmol) andtert-butyldimethylsilyl chloride (4.65 g, 30.86 mmol) were added and thereaction mixture was stirred overnight at room temperature. The reactionmixture was filtered to remove solids and washed with brine twice. Theorganic layer was dried over magnesium sulfate, filtered andconcentrated. Silica gel chromatography (0%-10% ethyl acetate/hexanes)afforded the title compound as a colorless oil (6.77 g, 64%).

MS: 342 ES+ (C₁₈H₃₅NO₃Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.04 (s, 6H); 0.39 (m, 2H); 0.63 (m, 2H);0.85 (s, 9H); 1.32 (m, 1H); 1.37 (m, 9H); 3.55 (m, 1H); 3.67 (m, 1H);3.99 (m, 1H); 4.63 (s, 1H); 4.78 (s, 1H); 6.80 (m, 1H).

Intermediate 234:(S)-1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-amine

To a solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-ylcarbamate(Intermediate 233, 6.77 g, 19.82 mmol) in DCM (100 mL) at roomtemperature was added zinc bromide (17.86 g, 79.28 mmol). The reactionmixture was stirred overnight at room temperature. Another 1 eq of zincbromide was added. After several hours the reaction mixture was filteredand washed with saturated sodium bicarbonate. The resulting emulsion wasfiltered through a nylon filter and the layers were separated. Theorganics were dried over magnesium sulfate, filtered and concentrated toafford the title compound as a yellow oil (4.61 g, 96%).

¹H NMR (300 MHz, DMSO-d₆) δ: 0.04 (s, 6H); 0.39 (m, 2H); 0.63 (m, 2H);0.87 (s, 9H); 1.35 (m, 1H); 1.81 (m, 2H); 3.33 (m, 1H); 3.45 (m, 1H);3.67 (m, 1H); 4.59 (s, 1H); 4.83 (m, 1H).

Intermediate 235: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate

The title compound was prepared from(S)-1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-amine(Intermediate 234, 4.61 g, 19.09 mmol) and2-bromo-N-methoxy-N-methylacetamide (Intermediate 4, 3.16 g, 17.36 mmol)following the procedure described for Intermediate 5. The desiredproduct was obtained as a light yellow oil (4.94 g, 64%).

MS: 443 ES+ (C₂₂H₄₂N₂O₅Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (m, 6H); 0.35 (m, 1H); 0.48 (m, 1H);0.61 (m, 2H); 0.83 (m, 9H); 1.35 (m, 9H); 3.07 (m, 3H); 3.65 (m, 3H);3.84 (m, 2H); 4.02 (m, 2H); 4.54 (m, 1H); 4.83 (m, 2H).

Intermediate 236: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-yl(2-oxopent-3-enyl)carbamate

A suspension of cerium (III) chloride (27.8 g, 112.95 mmol) in THF (100mL) at room temperature was stirred vigorously for 2 hours. Thesuspension was cooled to −78° C. and (E)-prop-1-enylmagnesium bromide(0.5 M in THF) (226 mL, 112.95 mmol) was added dropwise. The mixture wasstirred at −78° C. for 1.5 hours. (S)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 235, 5 g, 11.30 mmol) in THF (20 mL) was then addeddropwise at −78° C. The reaction was stirred at −78° C. for 30 minutesand then warmed to 0° C. for 15 minutes. The reaction was quenched with10% citric acid, diluted further with water and extracted twice withether. The organics were washed once with brine, dried over magnesiumsulfate, filtered and concentrated. Silica gel chromatography (0%-20%ethyl acetate/hexanes) afforded the title compound as a light yellow oil(4.0 g, 84%).

MS: 424 ES+ (C₂₃H₄₁Na₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.03 (m, 6H); 0.43 (m, 2H); 0.61 (m, 2H);0.83 (m, 9H); 1.34 (m, 10H); 1.84 (m, 2H); 2.04 (m, 1H); 3.74 (m, 1H);3.84 (m, 2H); 4.03 (m, 1H); 4.57 (m, 1H); 4.79 (m, 2H); 6.28 (m, 1H);6.84 (m, 1H).

Intermediate 237: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-cyclopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S,E)-tert-butyl1-(tert-butyldimethylsilyloxy)-3-cyclopropylbut-3-en-2-yl(2-oxopent-3-enyl)carbamate(Intermediate 236, 4 g, 9.44 mmol) following the procedure described forIntermediate 7, except the reaction mixture was heated at 110° C.overnight. The desired product was obtained as a light brown oil (2.97g, 82%).

MS: 382 ES+ (C₂₀H₃₅NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (m, 6H); 0.62 (m, 1H); 0.80 (s, 9H);1.00 (m, 3H); 1.42 (s, 9H); 1.61 (m, 1H); 3.80 (m, 1H); 3.95 (m, 2H);4.19 (m, 1H); 4.75 (m, 1H); 5.72 (s, 1H).

Intermediate 238: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-cyclopropyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-cyclopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 237, 2.97 g, 7.78 mmol) following the procedure describedfor Intermediate 8. The desired product was obtained as a tan oil (2.74g, 92%).

MS: 384 ES+ (C₂₀H₃₇NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (m, 6H); 0.34 (m, 1H); 0.47 (m, 1H);0.64 (m, 2H); 0.85 (m, 9H); 1.26 (m, 1H); 1.39 (s, 9H); 2.65 (m, 1H);3.89 (m, 3H); 4.05 (m, 1H); 4.95 (m, 1H); 5.34 (m, 1H).

Intermediate 239: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-3-cyclopropyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 238, 2.74 g, 7.14 mmol) andN-(allyloxy)-2-nitrobenzenesulfonamide (Intermediate 9, 1.85 g, 7.14mmol) following the procedure described for Intermediate 10. The desiredproduct was obtained as a light yellow oil (3.19 g, 71%).

MS: 624 ES+ (C₂₉H₄₅N₃O₈SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.00 (m, 6H); 0.34 (m, 1H); 0.63 (m, 2H);0.83 (m, 9H); 1.37 (m, 9H); 3.30 (m, 1H); 3.84 (m, 2H); 4.30 (m, 4H);5.18 (m, 2H); 5.75 (m, 1H); 8.04 (m, 4H).

Intermediate 240: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-cyclopropyl-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-3-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 239, 3.19 g, 5.11 mmol) following the procedure describedfor Intermediate 18. The desired product was obtained as a tan foam(2.35 g, 90%).

MS: 510 ES+ (C₂₃H₃₁H₃O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.32 (m, 2H); 0.62 (m, 2H); 1.35 (m, 9H);3.30 (m, 1H); 3.67 (m, 2H); 4.27 (m, 4H); 4.71 (m, 1H); 5.19 (m, 2H);5.71 (m, 1H); 8.04 (m, 4H).

Intermediate 241:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-cyclopropyl-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 240, 2.35 g, 4.61 mmol) following the procedure describedfor Intermediate 19. The desired product was obtained as an orange foam(2.28 g, 94%).

MS: 524 ES+ (C₂₃H₂₉N₃O₉S)

Intermediate 242: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 241, 2.28 g, 4.35 mmol) following the proceduredescribed for Intermediate 20. The desired product was obtained as anorange foam (1.07 g, 47%).

MS: 523 ES+ (C₂₃H₃₀N₄O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.23 (m, 2H); 0.59 (m, 2H); 1.35 (m, 9H);3.58 (m, 1H); 4.23 (m, 3H); 4.72 (m, 1H); 5.19 (m, 2H); 5.71 (m, 1H);7.18 (m, 1H); 7.59 (m, 1H); 8.04 (m, 4H).

Intermediate 243:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-3-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 242, 0.932 g, 1.78 mmol) following the procedure describedfor Intermediate 21. The desired product was obtained as an orange foam(0.518 g, 68%).

MS: 423 ES+ (C₁₈H₂₂N₄O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.18 (m, 2H); 0.53 (m, 2H); 1.29 (m, 1H);2.30 (m, 1H); 2.58 (m, 1H); 2.95 (m, 1H); 3.72 (m, 1H); 4.22 (m, 1H);4.36 (m, 2H); 4.96 (m, 1H); 5.24 (m, 2H); 5.80 (m, 1H); 7.07 (bs, 1H);7.39 (bs, 1H); 8.04 (m, 4H).

Intermediate 244:(R)-5-(allyloxyamino)-3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 243, 0.518 g, 1.23 mmol) following the procedure describedfor Intermediate 12. The desired product was obtained as a light yellowoil (0.171 g, 59%). The product is a mixture of diastereomers.

MS: 238 ES+ (C₁₂H₁₉N₃O₂)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.28 (m, 2H); 0.41 (m, 2H); 0.54 (m, 2H);1.33 (m, 1H); 2.49 (m, 1H); 2.64 (m, 1H); 2.93 (m, 1H); 3.23 (m, 1H);3.65 (m, 1H); 4.07 (m, 2H); 5.19 (m, 3H); 5.89 (m, 1H); 6.26 (m, 1H);6.97 (bs, 1H); 7.34 (bs, 1H).

Intermediate 245:(2S,5R)-6-(allyloxy)-3-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

The title compound was prepared from(R)-5-(allyloxyamino)-3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 244, 0.316 g, 1.33 mmol) following the procedure describedfor Intermediate 16. The desired product was obtained as a colorless oil(0.261 g, 74%).

MS: 264 ES+ (C₁₃H₁₇N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.37 (m, 2H); 0.60 (m, 2H); 1.20 (m, 1H);2.98 (m, 1H); 3.79 (m, 1H); 3.92 (m, 1H); 4.20 (m, 1H); 4.33 (m, 2H);5.28 (m, 2H); 5.93 (m, 2H); 7.30 (bs, 1H); 7.86 (bs, 1H).

Intermediate 246: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-3-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 245, 0.261 g, 0.99 mmol) following the procedure describedfor Intermediate 17. The desired product was obtained as a yellow foam(0.39 g, 65%).

MS: 302 ES−, 303 ES+ (C₁₀H₁₂N₃O₆S−.C₂₁H₂₀P+)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.39 (m, 2H); 0.58 (m, 2H); 1.18 (m, 2H);2.16 (m, 3H); 3.03 (m, 1H); 3.78 (m, 1H); 4.01 (m, 1H); 4.20 (m, 1H);5.91 (m, 1H); 6.65 (m, 1H); 7.38 (m, 2H); 7.78 (m, 15H).

Example 25(2S,5R)-4-(2-acetamidoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate Sodium Salt

The title compound was prepared from pyridine(2S,5R)-4-(2-acetamidoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 248, 0.007 g, 0.020 mmol) according to theprocedure described for Example 1. The desired product was obtained as awhite solid (3 mg, 58%).

MS: 347 ES− (C₁H₁₆N₄O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.85 (d, J=6.78 Hz, 1H) 1.70-1.84 (m, 3H)2.18 (t, J=6.78 Hz, 1H) 3.12-3.21 (m, 2H) 3.97-4.17 (m, 2H) 5.42 (br.s., 1H) 7.27 (br. s., 1H) 7.49 (s, 1H) 7.71 (s, 1H).

The Intermediates for Example 25 were Prepared as Follows Intermediate247:(2S,5R)-4-(2-acetamidoethyl)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of(2S,5R)-6-(benzyloxy)-4-(2-nitroethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 226, 0.300 g, 0.87 mmol) in ethanol (10 mL) was added zincdust (1.416 g, 21.66 mmol) and acetic acid (1.984 mL, 34.65 mmol). Thereaction mixture was stirred at ambient temperature for 1 hour. ThenDIPEA (2.118 mL, 12.13 mmol) was added to reaction mixture followed byacetic anhydride (0.245 mL, 2.60 mmol). The reaction mixture was stirredfor an additional 2 hours, then diluted with DCM, washed with saturatedbicarbonate and concentrated to dryness. Flash chromatography, 0%-10%MeOH/DCM, afforded the title compound as a clear oil (0.31 g, 61%).

MS: 359 ES+ (C₁₈H₂₂N₄O₄)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.75 (s, 3H) 2.08-2.21 (m, 2H) 2.92-3.05(m, 1H) 3.09-3.24 (m, 3H) 3.72 (s, 1H) 4.12 (br. s., 1H) 4.89 (s, 2H)5.44 (br. s., 1H) 7.29 (br. s., 1H) 7.34-7.51 (m, 6H) 7.70 (br. s., 1H).

Intermediate 248:(2S,5R)-4-(2-acetamidoethyl)-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of4-(2-acetamidoethyl)-6-(benzyloxy)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 247, 0.120 g, 0.33 mmol) in EtOAc (20 mL) and ethanol(6.67 mL) was added Pd/C (0.036 g, 0.33 mmol). The reaction mixture wasstirred under hydrogen at 1 atm for 1 hour, then filtered through celiteand concentrated to dryness to afford the title compound as a whitesolid (0.09 g, 100%).

Intermediate 249: pyridine(2S,5R)-4-(2-acetamidoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

To a solution of(2S,5R)-4-(2-acetamidoethyl)-6-hydroxy-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 248, 0.090 g, 0.34 mmol) in pyridine (5 mL) was addedpyridine/sulfur trioxide complex (0.320 g, 2.01 mmol). The reactionmixture was stirred overnight at room temperature. The solvent wasremoved in vacuo and the residue was purified by reverse phase HPLC. Thetitle compound was obtained as a white solid after lypholization (0.007g, 6%).

MS: 347 ES− (C₁₁H₁₆N₄O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.85 (d, J=6.78 Hz, 1H) 1.68-1.82 (m, 3H)2.18 (t, J=7.16 Hz, 2H) 3.06 (dt, J=12.81, 6.40 Hz, 1H) 3.47 (d, J=6.78Hz, 1H) 3.99-4.17 (m, 3H) 5.42 (d, J=1.51 Hz, 1H) 7.28 (br. s., 1H) 7.49(br. s., 1H) 7.72 (t, J=5.27 Hz, 1H) 7.91-8.02 (m, 1H) 8.48 (t, J=7.91Hz, 1H) 8.89 (br. s., 1H).

Example 26(2S,5R)-2-(methoxymethyl)-4-(methylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound (0.08 g, 44%) was prepared according to the proceduredescribed for Example 1, starting from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(methoxymethyl)-4-(methylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 260, 0.33 g).

Optical rotation: (0.1 g/dL, MeOH)=−136

MS: 320 ES− (C₁₀H₁₅N₃O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.65 (d, J=4.52 Hz, 3H) 3.19-3.29 (m, 6H)3.53-3.69 (m, 2H) 3.89 (td, J=5.75, 3.01 Hz, 1H) 4.50-4.61 (m, 1H) 6.38(dd, J=3.01, 0.94 Hz, 1H) 7.76 (d, J=4.52 Hz, 1H).

The Intermediates for Example 26 were Prepared as Follows Intermediate250: (S)-tert-butyl1-hydroxybut-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate

To a solution of (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 5, 10.7 g, 26.58 mmol) in THF (25 mL) was added TBAF (31.9mL, 31.89 mmol) at 0° C. After 1 hour the reaction mixture wasconcentrated and purified by silica gel chromatography (hexanes/ethylacetate) to afford the product as a light yellow oil (5.9 g, 77%).

MS: 289 ES+ (C₁₃H₂₄N₂O₅)

¹H NMR (300 MHz, CDCl₃) δ: 1.36-1.61 (m, 10H) 3.25 (s, 3H) 3.35-3.52 (m,1H) 3.57-3.72 (m, 2H) 3.72-3.82 (m, 3H) 4.26-4.56 (m, 1H) 4.83 (ddd,J=8.24, 4.19, 2.07 Hz, 2H) 5.10-5.34 (m, 2H) 5.57-5.84 (m, 1H).

Intermediate 251: (S)-tert-butyl2-(methoxy(methyl)amino)-2-oxoethyl(1-methoxybut-3-en-2-yl)carbamate

To a solution of (S)-tert-butyl1-hydroxybut-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 250, 7 g, 24.28 mmol) in THF (75 mL) at 0° C. was addeddimethyl sulfate (2.436 mL, 25.49 mmol) followed by LiHMDS (26.7 mL,26.70 mmol). The reaction mixture was allowed to warm slowly to roomtemperature and stirred for two hours. The reaction mixture was thenpartitioned between ethyl acetate and water. The layers were separatedand the organics were washed with saturated sodium bicarbonate, waterand brine. The organics were dried over magnesium sulfate, filtered andconcentrated. Silica gel chromatography (5%-30% ethyl acetate/hexanes)afforded the title compound as a clear oil (6.89 g, 91%).

MS: 303 ES+ (C₁₄H₂₆₄N₂O₅)

¹H NMR (300 MHz, CDCl₃) δ: 1.18-1.42 (m, 9H) 3.02 (s, 3H) 3.14 (d,J=6.03 Hz, 3H) 3.41-3.64 (m, 5H) 3.76-4.13 (m, 2H) 4.57-4.82 (m, 1H)4.93-5.13 (m, 2H) 5.60-5.90 (m, 1H).

Intermediate 252: (S)-tert-butyl1-methoxybut-3-en-2-yl(2-oxoethyl)carbamate

To a solution of (S)-tert-butyl2-(methoxy(methyl)amino)-2-oxoethyl(1-methoxybut-3-en-2-yl)carbamate(Intermediate 251, 3.35 g, 11.08 mmol) in dichloromethane (60 ml) at−78° C. under nitrogen was added diisobutylaluminum hydride (1.5 eq,1.0M in dichloromethane) (16.62 ml, 16.62 mmol). The reaction mixturewas stirred at that temperature for two hours. The reaction was quenchedwith slow addition of methanol at the low temperature and slowly warmedto ambient temperature. The reaction mixture was diluted withdichloromethane and washed with 10% aqueous potassium sodium tartratetwice, followed by water and brine. The organics were dried over sodiumsulfate, filtered and concentrated. Silica gel chromatography (0%-50%ethyl acetate/hexanes) afforded the title compound as a clear oil (1.8g, 68%).

¹H NMR (300 MHz, CDCl₃) δ: 1.46 (s, 9H) 3.31 (s, 3H) 3.46-3.65 (m, 2H)3.65-3.80 (m, 1H) 3.80-4.05 (m, 1H) 4.65-5.01 (m, 1H) 5.13-5.36 (m, 2H)5.64-5.90 (m, 1H) 9.49 (s, 1H).

Intermediate 253: methyl4-(tert-butoxycarbonyl((S)-1-methoxybut-3-en-2-yl)amino)-3-hydroxy-2-methylenebutanoate

To a solution of (S)-tert-butyl1-methoxybut-3-en-2-yl(2-oxoethyl)carbamate (Intermediate 252, 1.475 g,6.06 mmol) and methyl acrylate (0.82 ml, 9.09 mmol) in MeOH (0.2 ml)under nitrogen at room temperature was added quinuclidine (0.337 g, 3.03mmol). The reaction mixture was stirred at that temperature overnight.Another 100 mg of quinuclidine was added and the reaction was stirredfor another 24 hours. The reaction mixture was then partitioned betweenwater and ethyl acetate. The layers were separated and the organics werewashed with water and brine, then dried over magnesium sulfate, filteredand concentrated. Silica gel chromatography (0%-30% ethylacetate/hexanes) afforded the title compound as a clear oil (1.28 g,65%). The product is a mixture of two diastereomers.

MS: 330 ES+ (C₁₆H₂₇NO₆)

Intermediate 254: (2S,5S)-1-tert-butyl 4-methyl5-hydroxy-2-(methoxymethyl)-5,6-dihydropyridine-1,4(2H)-dicarboxylate

The title compound (0.45 g, 40%) was prepared according to the proceduredescribed for Intermediate 101 starting from methyl4-(tert-butoxycarbonyl((S)-1-methoxybut-3-en-2-yl)amino)-3-hydroxy-2-methylenebutanoate(Intermediate 253, 1.22 g).

MS: 302 ES+ (C₁₄H₂₃NO₆)

¹H NMR (300 MHz, CDCl₃) δ: 1.49 (s, 9H) 2.96-3.18 (m, 1H) 3.39 (s, 3H)3.52-3.73 (m, 2H) 3.88 (s, 3H) 3.99-4.19 (m, 1H) 4.19-4.38 (m, 1H)4.52-4.59 (m, 1H) 4.59-4.81 (m, 1H) 7.01 (d, J=4.14 Hz, 1H).

Intermediate 255: (2S,5S)-tert-Butyl5-hydroxy-2-(methoxymethyl)-4-(methylcarbamoyl)-5,6-dihydropyridine-1(2H)-carboxylate

A dry 2 necked round bottomed flask was equipped with a magnetic stirrerand it was flushed with nitrogen twice. Then trimethylaluminum (0.191ml, 0.38 mmol) was added followed by toluene (0.5 ml). The solution wascooled down to −10° C. and methanamine (0.183 ml, 0.37 mmol) was addedslowly. The reaction mixture was stirred at that temperature for 20minutes. A solution of (2S,5S)-1-tert-butyl 4-methyl5-hydroxy-2-(methoxymethyl)-5,6-dihydropyridine-1,4(2H)-dicarboxylate(Intermediate 254, 100 mg, 0.33 mmol) in toluene (0.5 ml) was addedslowly at room temperature. After stirring overnight the reaction wasquenched with 1M HCl and allowed to stir for 15 minutes to complete thehydrolysis. The reaction mixture was extracted with ethyl acetate. Theorganics were washed with saturated sodium bicarbonate, dried overmagnesium sulfate, filtered and concentrated. Silica gel chromatography(MeOH/DCM) afforded the title compound as a clear oil (53 mg).

MS: 302 ES+ (C₁₄H₂₄N₂O₅)

¹H NMR (300 MHz, CDCl₃) δ: 1.48 (s, 9H) 1.69 (br. s., 2H) 2.57-2.94 (m,3H) 3.08 (dd, J=13.00, 9.04 Hz, 1H) 3.36 (s, 3H) 3.40-3.56 (m, 1H)3.51-3.72 (m, 1H) 4.13 (dd, J=13.00, 4.71 Hz, 1H) 4.47 (t, J=6.78 Hz,1H) 4.60-4.66 (m, 1H) 6.60 (d, J=4.14 Hz, 1H).

Intermediate 256: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(methoxymethyl)-4-(methylcarbamoyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound, as an off-white solid (1 g, 63%), was preparedaccording to the procedure for Intermediate 10 starting from(2S,5S)-tert-Butyl5-hydroxy-2-(methoxymethyl)-4-(methylcarbamoyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 255, 0.87 g).

MS: 541 ES+ (C₂₃H₃₂N₄O₉S)

Intermediate 257:(3R,6S)-3-(N-(allyloxy)-2-nitrophenylsulfonamido)-6-(methoxymethyl)-N-methyl-1,2,3,6-tetrahydropyridine-4-carboxamide

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-(methoxymethyl)-4-(methylcarbamoyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 256, 1 g, 1.85 mmol) in dioxane (10 ml) at roomtemperature was added 4M HCl in Dioxane (2 ml, 57.16 mmol). The reactionmixture was stirred at that temperature for 4-5 hours. The reactionmixture was concentrated and the crude was taken up in 2M NaOH andextracted with dichloromethane. The combined organic extracts werewashed with brine, dried over sodium sulfate, filtered and concentrated.Silica gel chromatography (increasing percentage of methanol indichloromethane) afforded the title compound as a thick oil (0.56 g,69%).

MS: 441 ES+ (C₁₈H₂₄N₄O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.58 (d, J=3.96 Hz, 3H); 2.77 (br. s., 1H);3.14-3.26 (m, 2H); 3.24 (s, 3H) 3.48 (d, J=4.52 Hz, 1H) 4.16-4.42 (m,2H) 4.66 (br. s., 1H) 5.12-5.33 (m, 2H) 5.83 (dd, J=17.24, 10.46 Hz, 1H)6.63 (d, J=2.26 Hz, 1H) 7.77-7.94 (m, 2H) 7.94-8.02 (m, 2H) 8.08 (d,J=7.91 Hz, 1H) (2H peaks buried underneath water peak).

Intermediate 258:(3R,6S)-3-(allyloxyamino)-6-(methoxymethyl)-N-methyl-1,2,3,6-tetrahydropyridine-4-carboxamide

To a mixture of(3R,6S)-3-(N-(allyloxy)-2-nitrophenylsulfonamido)-6-(methoxymethyl)-N-methyl-1,2,3,6-tetrahydropyridine-4-carboxamide(Intermediate 257, 561 mg, 1.27 mmol) and potassium carbonate (880 mg,6.37 mmol) in acetonitrile (15 mL) at room temperature was addedbenzenethiol (0.654 mL, 6.37 mmol). The resulting mixture was stirred atroom temperature overnight. The reaction mixture was concentrated ontosilica gel. Silica gel chromatography (0%-20% methanol/dichloromethane)afforded the title compound as an off-white solid (0.26 g, 81%).

MS: 256 ES+ (C₁₂H₂₁N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.64 (d, J=4.71 Hz, 3H) 2.73 (d, J=9.04 Hz,1H) 2.94 (dd, J=12.90, 3.11 Hz, 1H) 3.20-3.28 (m, 2H) 3.26 (s, 3H)3.37-3.52 (m, 1H) 3.54-3.68 (m, 1H) 4.12 (dt, J=5.70, 1.39 Hz, 2H)5.00-5.30 (m, 2H) 5.79-6.04 (m, 1H) 6.43 (d, J=8.48 Hz, 1H) 6.53-6.65(m, 1H) 7.86 (d, J=4.52 Hz, 1H) (1H peak buried underneath water peak).

Intermediate 259:(2S,5R)-6-(allyloxy)-2-(methoxymethyl)-N-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide

The title compound (0.28 g, 68%) was prepared according to the proceduredescribed for Intermediate 13, starting from(3R,6S)-3-(allyloxyamino)-6-(methoxymethyl)-N-methyl-1,2,3,6-tetrahydropyridine-4-carboxamide(Intermediate 258, 0.52 g)

MS: 282 ES+ (C₁₃H₁₉N₃O₄)

¹H NMR (300 MHz, CDCl₃) δ: 2.88 (d, J=4.90 Hz, 3H) 3.27-3.38 (m, 2H)3.40 (s, 3H) 3.49-3.79 (m, 2H) 4.11 (td, J=5.79, 2.92 Hz, 1H) 4.36-4.41(m, 1H) 4.41-4.56 (m, 2H) 5.14-5.46 (m, 2H) 5.94 (br. s., 1H) 5.96-6.14(m, 1H) 6.26 (d, J=2.26 Hz, 1H).

Intermediate 260: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-(methoxymethyl)-4-(methylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound (0.33 g, 79%) was prepared according to the proceduredescribed for Intermediate 17, starting from(2S,5R)-6-(allyloxy)-2-(methoxymethyl)-N-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-4-carboxamide(Intermediate 259, 0.19 g).

MS: 319 ES−, 303 ES+ (C₁₀H₁₅N₃O₇S, C₂₁H₂₀P)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.17 (dt, J=6.36, 2.00 Hz, 4H) 2.65 (d,J=4.71 Hz, 3H) 3.17-3.28 (m, 4H) 3.51-3.69 (m, 2H) 3.89 (td, J=5.79,3.11 Hz, 1H) 4.57 (d, J=1.13 Hz, 1H) 6.38 (dd, J=2.92, 1.04 Hz, 1H)6.54-6.78 (m, 1H) 7.18-7.41 (m, 1H) 7.63-7.84 (m, 14H) 7.85-8.00 (m,3H).

Example 27(2S,5R)-2-carbamoyl-4-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 272, 0.305 g, 0.50 mmol) following the proceduredescribed for Example 1. The desired product was obtained as a whitesolid (66.4 mg, 40%) after HPLC purification (Synergi Polar RP 19 mm×100mm 5 μm).

Optical rotation: (0.1 g/dL, MeOH)=−229

MS: 302 ES− (C₁₀H₁₃N₃O₆S)

¹H NMR (300 MHz, D₂O) δ: 0.56 (m, 1H); 0.78 (m, 3H); 1.56 (m, 1H); 3.28(d, 1H); 3.59 (m, 1H); 4.07 (m, 1H); 4.51 (m, 1H); 5.63 (m, 1H).

The Intermediates for Example 27 were Prepared as Follows Intermediate261:(E)-N′-(1-cyclopropylethylidene)-2,4,6-triisopropylbenzenesulfonohydrazide

The title compound was prepared from 2,4,6-triisopropylbenzenesulfonylhydrazide (Aldrich, 20 g, 67.01 mmol) and 1-cyclopropylethanone(Aldrich, 6.28 mL, 67.01 mmol) following the procedure described forIntermediate 33. The desired product was obtained as a white solid (15g, 61%).

MS: 365 ES+ (C₂₀H₃₂N₂O₂S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.56 (m, 3H); 0.75 (m, 1H); 1.17 (m, 18H);1.47 (m, 1H); 1.61 (s, 3H); 2.91 (m, 1H); 4.25 (m, 2H); 7.20 (s, 2H);9.97 (s, 1H).

Intermediate 262: (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(3-cyclopropyl-2-oxobut-3-enyl)carbamate

The title compound was prepared from(E)-N′-(1-cyclopropylethylidene)-2,4,6-triisopropylbenzenesulfonohydrazide(Intermediate 261, 17.58 g, 48.22 mmol) and (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(2-(methoxy(methyl)amino)-2-oxoethyl)carbamate(Intermediate 5, 9.71 g, 24.11 mmol) following the procedure describedfor Intermediate 34. The desired product was obtained as a light yellowoil (4.86 g, 49%).

MS: 410 ES+ (C₂₂H₃₉NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (m, 6H); 0.45 (m, 2H); 0.75 (m, 2H);0.83 (m, 10H); 1.32 (m, 9H); 0.70 (m, 1H); 3.71 (m, 2H); 4.35 (m, 2H);5.18 (m, 2H); 5.46 (m, 1H); 5.78 (m, 1H); 5.88 (m, 1H).

Intermediate 263: (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-cyclopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl1-(tert-butyldimethylsilyloxy)but-3-en-2-yl(3-cyclopropyl-2-oxobut-3-enyl)carbamate(Intermediate 262, 5.16 g, 12.60 mmol) following the procedure describedfor Intermediate 7, except the reaction mixture was heated at 85° C.overnight The desired product was obtained as a light yellow oil (3.82g, 79%).

MS: 382 ES+ (C₂₀H₃₅NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (m, 6H); 0.45 (m, 2H); 0.75 (m, 1H);0.83 (m, 10H); 1.28 (m, 1H); 1.41 (m, 9H); 1.71 (m, 1H); 3.85 (m, 2H);4.05 (m, 1H); 4.66 (m, 1H); 6.56 (m, 1H).

Intermediate 264: (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-cyclopropyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-cyclopropyl-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 263, 3.82 g, 10.01 mmol) following the procedure describedfor Intermediate 8. The desired product was obtained as a colorless oil(3 g, 78%).

MS: 384 ES+ (C₂₀H₃₇NO₄Si)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.02 (s, 6H); 0.22 (m, 1H); 0.57 (m, 3H);0.85 (s, 9H); 1.38 (s, 9H); 1.57 (m, 1H); 2.69 (m, 1H); 3.58 (m, 2H);3.93 (m, 1H); 4.17 (m, 1H); 5.11 (d, 1H); 5.22 (m, 1H).

Intermediate 265: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5S)-tert-butyl2-((tert-butyldimethylsilyloxy)methyl)-4-cyclopropyl-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 264, 3 g, 7.82 mmol) andN-(allyloxy)-2-nitrobenzenesulfonamide (Intermediate 9, 2.020 g, 7.82mmol) following the procedure described for Intermediate 10. The desiredproduct was obtained as a yellow oil (3.62 g, 74%).

MS: 624 ES+ (C₂₉H₄₅N₃O₈SSi)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.01 (s, 6H); 0.40 (m, 3H); 0.83 (s, 9H);1.31 (m, 9H); 3.14 (m, 1H); 3.57 (m, 2H); 4.10 (m, 4H); 4.43 (m, 1H);5.20 (m, 2H); 5.70 (m, 2H); 8.02 (m, 4H).

Intermediate 266: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-cyclopropyl-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-((tert-butyldimethylsilyloxy)methyl)-4-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 265, 3.62 g, 5.80 mmol) in THF (30 mL) following theprocedure described for Intermediate 18. The desired product wasobtained as an off-white foam (2.64 g, 89%).

MS: 510 ES+ (C₂₃H₃₁N₃O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.46 (m, 3H); 1.34 (m, 9H); 3.07 (m, 1H);3.38 (m, 2H); 4.05 (m, 2H); 4.29 (m, 3H); 4.73 (m, 1H); 5.20 (m, 2H);5.69 (m, 2H); 8.04 (m, 4H).

Intermediate 267:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-cyclopropyl-2-(hydroxymethyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 266, 2.64 g, 5.18 mmol) following the procedure describedfor Intermediate 19. The desired product was obtained as an orange foam(2.33 g, 86%).

MS: 524 ES+ (C₂₃H₂₉N₃O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.46 (m, 3H); 1.28 (m, 9H); 3.10 (m, 1H);3.90 (m, 1H); 4.29 (m, 3H); 4.78 (m, 1H); 5.19 (m, 2H); 5.69 (m, 2H);8.04 (m, 4H).

Intermediate 268: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-1-(tert-butoxycarbonyl)-4-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxylicacid (Intermediate 267, 2.33 g, 4.45 mmol) following the proceduredescribed for Intermediate 20. The desired product was obtained as a tanfoam (1.47 g, 63%).

MS: 523 ES+ (C₂₃H₃₀N₄O₈S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.53 (m, 4H); 1.28 (m, 9H); 3.15 (m, 1H);3.93 (m, 1H); 4.27 (m, 3H); 4.73 (m, 1H); 5.19 (m, 2H); 5.73 (m, 2H);7.03 (m, 1H); 7.43 (m, 1H); 8.05 (m, 4H).

Intermediate 269:(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-2-carbamoyl-4-cyclopropyl-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 268, 1.47 g, 2.81 mmol) following the procedure describedfor Intermediate 21. The desired product was obtained as a light yellowfoam (0.95 g, 80%).

MS: 423 ES+ (C₁₈H₂₂N₄O₆S)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.30 (m, 1H); 0.50 (m, 2H); 0.81 (m, 1H);1.28 (m, 1H); 2.70 (m, 2H); 3.75 (m, 1H); 4.07 (m, 1H); 4.37 (m, 2H);5.24 (m, 2H); 5.84 (m, 2H); 7.02 (m, 1H); 7.28 (m, 1H); 8.04 (m, 4H).

Intermediate 270:(R)-5-(allyloxyamino)-4-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from(2S,5R)-5-(N-(allyloxy)-2-nitrophenylsulfonamido)-4-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 269, 0.95 g, 2.25 mmol) following the procedure describedfor Intermediate 22. The desired product was obtained as a light yellowoil (0.307 g, 57%).

MS: 238 ES+ (C₁₂H₁₉N₃O₂)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.52 (m, 4H); 1.43 (m, 2H); 2.46 (m, 1H);3.04 (m, 2H); 3.59 (m, 1H); 4.13 (m, 2H); 5.20 (m, 2H); 5.39 (m, 1H);5.92 (m, 1H); 6.39 (m, 1H); 7.00 (m, 1H); 7.34 (bs, 1H).

Intermediate 271:(2S,5R)-6-(allyloxy)-4-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

The title compound was prepared from(R)-5-(allyloxyamino)-4-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide(Intermediate 270, 0.307 g, 1.29 mmol) following the procedure describedfor Intermediate 16. The desired product was obtained as a yellow oil(0.168 g, 49%).

MS: 264 ES+ (C₁₃H₁₇N₃O₃)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.56 (m, 4H); 1.43 (m, 1H); 3.17 (m, 2H);3.71 (m, 1H); 4.11 (m, 1H); 4.36 (m, 2H); 5.34 (m, 3H); 5.94 (m, 1H);7.27 (bs, 1H); 7.49 (bs, 1H).

Intermediate 272: (E)-triphenyl(prop-1-enyl)phosphonium(2S,5R)-2-carbamoyl-4-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-4-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 271, 0.168 g, 0.64 mmol) following the procedure describedfor Intermediate 17. The desired product was obtained as a yellow foam(0.305 g, 79%).

MS: 302 ES−, 303 ES+ (C₁₀H₁₂N₃O₆S—.C₂₁H₂₀P+)

¹H NMR (300 MHz, DMSO-d₆) δ: 0.56 (m, 3H); 0.85 (m, 1H); 1.42 (m, 1H);2.16 (m, 3H); 3.18 (m, 2H); 3.71 (m, 1H); 4.10 (m, 1H); 5.44 (m, 1H);6.65 (m, 1H); 7.25 (m, 2H); 7.51 (m, 1H); 7.79 (m, 15H).

Example 28(2S,5R)-3-(2-methoxyethyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The title compound was prepared from(E)-triphenyl(prop-1-en-1-yl)phosphonium(2S,5R)-3-(2-methoxyethyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 288, 0.328 g, 0.53 mmol) following the proceduredescribed for Example 1. The desired product was obtained as anoff-white solid (146 mg, 81%).

Optical Rotation: (0.22 g/dL, DMSO)=−263

MS: 321 ES− (C₁₁H₁₈N₂O₇S)

¹H NMR (300 MHz, DMSO-d₆) δ: 2.13 (m, 2H); 3.07 (m, 1H); 3.22 (s, 3H);3.28 (s, 3H); 3.36 (m, 3H); 3.68 (m, 3H); 4.00 (m, 1H); 6.03 (m, 1H).

The Intermediates for Example 28 were Prepared as Follows Intermediate273: ((3-bromobut-3-en-1-yl)oxy)(tert-butyl)dimethylsilane

To a solution of 3-bromobut-3-en-1-ol (ACROS, 15.18 g, 100.53 mmol) inDCM (300 mL) at room temperature was added imidazole (8.90 g, 130.69mmol), 4-dimethylaminopyridine (2.456 g, 20.11 mmol) andtert-butyldimethylsilyl chloride (16.67 g, 110.58 mmol). The reactionmixture was stirred at room temperature for 3 hours, then filtered toremove solids and washed with brine. The organics were dried overmagnesium sulfate, filtered and concentrated. Silica gel chromatography(0%-15% ethyl acetate/hexanes) afforded the title compound (24.67 g,93%) as a colorless oil.

¹H NMR (300 MHz, CDCl₃) δ 0.08 (s, 3H), 0.90 (s, 9H), 2.63 (t, 2H), 3.80(t, 2H), 5.46 (m, 1H), 5.64 (m, 1H).

Intermediate 274:(S,E)-N-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-2-methylpropane-2-sulfinamide

To a solution of 2-((tert-butyldimethylsilyl)oxy)acetaldehyde (Aldrich,15 g, 86.05 mmol) in DCM (200 mL) at room temperature was addedcopper(II) sulfate (41.2 g, 258.16 mmol) and(S)-2-methylpropane-2-sulfinamide (Aldrich, 15.64 g, 129.08 mmol). Thereaction mixture was stirred overnight at room temperature, thenfiltered through celite, washed with DCM and concentrated to afford anoil. Silica gel chromatography (0%-25% ethyl acetate/hexanes) affordedthe title compound (14.73 g, 61.7%) as a colorless oil.

¹H NMR (300 MHz, CDCl₃) δ 0.11 (s, 6H), 0.93 (s, 9H), 1.22 (s, 9H), 4.55(d, 2H), 8.07 (t, 1H).

Intermediate 275:(S)-2-methyl-N—((S)-2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-yl)propane-2-sulfinamide

To a solution of ((3-bromobut-3-en-1-yl)oxy)(tert-butyl)dimethylsilane(Intermediate 273, 24.66 g, 92.97 mmol) in THF (200 mL) at −78° C. wasadded tert-butyllithium (1.7M in pentane) (120 mL, 204.54 mmol) dropwisevia cannula. The reaction mixture was stirred for 45 minutes at −78° C.The(S,E)-N-(2-((tert-butyldimethylsilyl)oxy)ethylidene)-2-methylpropane-2-sulfinamide(Intermediate 274, 17.2 g, 61.98 mmol) in THF (50 mL) was addeddropwise. The reaction mixture was stirred for ˜1.5 hours at −78° C. Thereaction was quenched with saturated sodium bicarbonate and extractedtwice with ether. The combined organic extracts were dried overmagnesium sulfate, filtered and concentrated. Silica gel chromatography(0%-15% ethyl acetate/hexanes) afforded the title compound (21.07 g,73.3%) as a colorless oil.

¹H NMR (300 MHz, CDCl₃) δ 0.06 (m, 12H), 0.90 (m, 18H), 1.24 (s, 9H),2.25 (m, 2H), 3.57 (m, 1H); 3.73 (m, 3H); 3.95 (m, 2H); 5.04 (m, 1H);5.19 (m, 1H).

Intermediate 276: (S)-2-amino-3-methylenepentane-1,5-diol hydrochloride

To a solution of2-methyl-N—((S)-2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-yl)propane-2-sulfinamide(Intermediate 275, 21.07 g, 45.42 mmol) in methanol (100 mL) at 0° C.was added hydrochloric acid (4M in dioxane) (22.71 mL, 90.85 mmol). Thereaction mixture was stirred at 0° C. for ˜20 minutes. LC/MS shows noremaining starting material. The reaction mixture was concentrated toafford an oil (7.6 g, 100%).

MS: 132 ES+ (C₆H₁₃NO₂)

Intermediate 277:(S)-2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-amine

To a solution of (S)-2-amino-3-methylenepentane-1,5-diol, HCl(Intermediate 276, 7.6 g, 45.34 mmol) in DCM (200 mL) at roomtemperature was added imidazole (12.35 g, 181.35 mmol),4-dimethylaminopyridine (2.77 g, 22.67 mmol) and tert-butyldimethylsilylchloride (20.50 g, 136.01 mmol). The reaction mixture was stirred atroom temperature overnight, then filtered to remove the solids andwashed with brine. The organics were dried over magnesium sulfate,filtered and concentrated. Silica gel chromatography (0%-5%methanol/dichloromethane) afforded the title compound (14.34 g, 88%) asa yellow oil.

MS: 359 ES+ (C₁₈H₄₁NO₂Si₂)

¹H NMR (300 MHz, CDCl₃) δ 0.06 (m, 12H), 0.90 (m, 18H), 2.30 (m, 2H),3.41 (m, 2H); 3.72 (m, 3H); 4.90 (m, 1H); 5.09 (m, 1H).

Intermediate 278: (S)-tert-butyl(2-(methoxy(methyl)amino)-2-oxoethyl)(2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-yl)carbamate

A mixture of(S)-2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-amine(14.34 g, 39.87 mmol) and potassium carbonate (Intermediate 277, 5.51 g,39.87 mmol) in DMF (300 mL) was stirred at room temperature for 1 hour.2-bromo-N-methoxy-N-methylacetamide (Intermediate 4, 7.26 g, 39.87 mmol)was added and the reaction mixture was stirred at room temperatureovernight. The reaction mixture was diluted with ethyl acetate andwashed with saturated sodium bicarbonate. An emulsion formed and wasfiltered to remove the solids. The layers were separated and theorganics were washed with 1:1 brine:water. The organics were dried overmagnesium sulfate, filtered and concentrated. The resulting yellow oilwas dissolved in THF (100 mL) and di-tert-butyl dicarbonate (17.40 g,79.73 mmol) was added. The reaction mixture was stirred at roomtemperature for 4 hours, then at 50° C. for ˜4 hours, then roomtemperature over the weekend. The reaction mixture was diluted withethyl acetate and washed with saturated sodium bicarbonate. The layerswere separated and the organics were washed twice with 1:1 brine:water,dried over magnesium sulfate, filtered and concentrated. Silica gelchromatography (0%-30% ethyl acetate/hexanes) afforded the titlecompound (13.55 g, 60.6%) as a light yellow oil.

MS: 561 ES+ (C₂₇H₅₆N₂O₆Si₂)

¹H NMR (300 MHz, CDCl₃) δ 0.06 (m, 12H), 0.88 (m, 18H), 1.45 (m, 9H),2.32 (m, 2H); 3.16 (s, 3H); 3.69 (m, 5H); 3.80 (m, 2H); 3.02 (m, 2H);4.64 (m, 1H); 5.04 (m, 2H).

Intermediate 279: (S)-tert-butyl(2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-yl)(2-oxopent-3-en-1-yl)carbamate

A suspension of cerium (III) chloride (47.6 g, 193.26 mmol) in THF (200mL) 3 at room temperature was stirred vigorously for 2 hours. Thesuspension was cooled to −78° C. and (E)-prop-1-enylmagnesium bromide(0.5 M in THF) (387 mL, 193.26 mmol) was added dropwise. The mixture wasstirred at −78° C. for 1.5 hours. (5)-tert-butyl(2-(methoxy(methyl)amino)-2-oxoethyl)(2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-yl)carbamate(Intermediate 278, 13.55 g, 24.16 mmol) in THF (50 mL) was then addeddropwise at −78° C. The reaction was stirred at −78° C. for 1 hour andthen warmed to 0° C. for 30 minutes. The reaction was quenched with 10%citric acid, diluted further with water and extracted twice with ether.The organics were dried over magnesium sulfate, filtered andconcentrated. Silica gel chromatography (0%-20% ethyl acetate/hexanes)afforded the title compound (10 g, 76%) as a yellow oil.

MS: 542 ES+ (C₂₈H₅₅NO₅Si₂)

¹H NMR (300 MHz, DMSO-d₆) δ 0.03 (m, 12H), 0.85 (m, 18H), 1.33 (m, 9H),1.84 (m, 2H); 2.18 (m, 2H); 3.71 (m, 6H); 4.56 (m, 1H); 4.93 (m, 2H);6.20 (m, 1H); 6.85 (m, 1H).

Intermediate 280: (S)-tert-butyl3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate

A solution of (S,E)-tert-butyl(2,2,3,3,11,11,12,12-octamethyl-7-methylene-4,10-dioxa-3,11-disilatridecan-6-yl)(2-oxopent-3-en-1-yl)carbamate(Intermediate 279, 10 g, 18.45 mmol) in toluene (400 mL) was purged withnitrogen for 15 minutes. The Hoveyda-Grubbs Catalyst 2nd Generation(2.320 g, 3.69 mmol) was then added. The reaction was heated at 100° C.overnight. Another 0.05 eq of catalyst was added and the reactionmixture was heated at 100° C. for another 2 hours. The reaction mixturewas concentrated onto silica gel. Silica gel chromatography (0%-15%ethyl acetate/hexanes) afforded the title compound (8.55 g, 93%) as alight brown oil.

MS: 500 ES+ (C₂₅H₄₉NO₅Si₂)

¹H NMR (300 MHz, DMSO-d₆) δ 0.01 (m, 12H), 0.81 (m, 18H), 1.42 (s, 9H),2.50 (m, 2H); 3.79 (m, 5H); 4.20 (m, 1H); 4.67 (m, 1H); 6.07 (s, 1H).

Intermediate 281: (2S,5S)-tert-butyl3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate

The title compound was prepared from (S)-tert-butyl3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5-oxo-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 280, 8.55 g, 17.11 mmol) according to the proceduredescribed for Intermediate 8 to afford the desired product (7.33 g, 85%)as a light yellow oil.

MS: 502 ES+ (C₂₅H₅₁NO₅Si₂)

¹H NMR (300 MHz, DMSO-d₆) δ 0.02 (m, 12H), 0.85 (m, 18H), 1.39 (s, 9H),2.21 (m, 2H); 2.68 (m, 1H); 3.70 (m, 4H); 4.02 (m, 2H); 4.21 (m, 1H);4.99 (m, 1H); 5.55 (s, 1H).

Intermediate 282: (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate

To a solution of (2S,5S)-tert-butyl3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-((tert-butyldimethylsilyl)oxy)methyl)-5-hydroxy-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 281, 7.33 g, 14.61 mmol) in toluene (100 mL) at roomtemperature was added triphenylphosphine (4.58 g, 17.53 mmol),N-(allyloxy)-2-nitrobenzenesulfonamide (3.77 g, 14.61 mmol) anddiisopropyl azodicarboxylate (3.45 mL, 17.53 mmol). The reaction mixturewas stirred overnight at room temperature then concentrated. Theresulting oil was triturated with hexane and filtered to removetriphenylphosphine oxide. The filtrate was concentrated onto silica gel.Silica gel chromatography (0%-20% ethyl acetate/hexanes) afforded thetitle compound (7.5 g, 69.2%) as a light yellow oil.

MS: 743 ES+ (C₃₄H₅₉N₃O₉SSi₂)

Intermediate 283:N-(allyloxy)-N-((3R,6S)-5-(2-((tert-butyldimethylsilyl)oxy)ethyl)-6-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)-2-nitrobenzenesulfonamide

To a solution of (2S,5R)-tert-butyl5-(N-(allyloxy)-2-nitrophenylsulfonamido)-3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-5,6-dihydropyridine-1(2H)-carboxylate(Intermediate 282, 7.5 g, 10.11 mmol) in DCM (100 mL) at roomtemperature was added zinc bromide (6.83 g, 30.32 mmol). The reactionmixture was stirred overnight at room temperature. Another equivalent ofzinc bromide was added and the reaction mixture was stirred for another24 hours. The reaction mixture was diluted with DCM and saturated sodiumbicarbonate was added. The biphasic mixture was filtered through celiteto remove the solids and the layers were separated. The organics werewashed once with brine, dried over magnesium sulfate, filtered andconcentrated to afford an orange oil (6.49 g, 100%).

MS: 642 ES+ (C₂₉H₅₁N₃O₇SSi₂)

Intermediate 284:O-allyl-N-((3R,6S)-5-(2-((tert-butyldimethylsilyl)oxy)ethyl)-6-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine

The title compound was prepared fromN-(allyloxy)-N-((3R,6S)-5-(2-((tert-butyldimethylsilyl)oxy)ethyl)-6-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)-2-nitrobenzenesulfonamide(Intermediate 283, 6.49 g, 10.11 mmol) according to the proceduredescribed for Intermediate 12. Silica gel chromatography (0%-5%methanol/dichloromethane) afforded the desired product (3.95 g, 86%) asa yellow oil.

MS: 457 ES+ (C₂₃H₄₈N₂O₃Si₂)

¹H NMR (300 MHz, DMSO-d₆) δ 0.03 (m, 12H), 0.85 (m, 18H), 2.15 (m, 2H),2.65 (m, 1H); 2.89 (m, 1H); 3.20 (m, 2H); 3.61 (m, 4H); 4.08 (m, 2H);5.15 (m, 2H); 5.44 (m, 1H); 5.88 (m, 1H); 6.16 (m, 1H).

Intermediate 285:(2S,5R)-6-(allyloxy)-3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a solution ofO-allyl-N-((3R)-5-(2-((tert-butyldimethylsilyl)oxy)ethyl)-6-(((tert-butyldimethylsilyl)oxy)methyl)-1,2,3,6-tetrahydropyridin-3-yl)hydroxylamine(Intermediate 284, 3.95 g, 8.65 mmol) and diisopropylethyl amine (6.02mL, 34.58 mmol) in acetonitrile (600 mL) at 0° C. was added triphosgene(0.871 g, 2.94 mmol) as a solution in acetonitrile (26 mL). Thetriphosgene solution was added at a rate of 4 mL/hour. Once addition wascomplete the reaction mixture was warmed to room temperature and stirredovernight. The reaction mixture was diluted with ethyl acetate, washedwith saturated sodium bicarbonate and 1:1 brine:water. The organics weredried over magnesium sulfate, filtered and concentrated. Silica gelchromatography (0%-30% ethyl acetate/hexanes) afforded the titlecompound as a yellow oil (3.69 g, 88%). This reaction was run in twoseparate 1 L flasks due to the large volume and combined for work up andpurification.

MS: 483 ES+ (C₂₄H₄₆N₂O₄Si₂)

¹H NMR (300 MHz, DMSO-d₆) δ 0.04 (m, 12H), 0.85 (m, 18H), 2.11 (m, 2H),3.01 (m, 1H); 3.44 (m, 1H); 3.63 (m, 3H); 3.93 (m, 2H); 4.32 (m, 2H);5.28 (m, 2H); 5.93 (m, 1H); 6.05 (m, 1H).

Intermediate 286:(2S,5R)-6-(allyloxy)-3-(2-hydroxyethyl)-2-(hydroxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a solution of(2S,5R)-6-(allyloxy)-3-(2-((tert-butyldimethylsilyl)oxy)ethyl)-2-(((tert-butyldimethylsilyl)oxy)methyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 285, 0.96 g, 1.99 mmol) in THF (6 mL) at 0° C. was addedtetrabutylammonium fluoride (1M in THF) (5.97 mL, 5.97 mmol). Thereaction mixture was stirred for ˜1 hour and then concentrated ontosilica gel. Silica gel chromatography (0%-10% methanol/dichloromethane)afforded the title compound (0.446 g, 88%) as a cloudy yellow oil.

MS: 255 ES+ (C₁₂H₁₈N₂O₄)

¹H NMR (300 MHz, DMSO-d₆) δ 2.06 (m, 2H), 2.98 (m, 1H), 3.42 (m, 3H),3.60 (m, 1H); 3.71 (m, 2H); 3.88 (m, 1H); 4.33 (m, 2H); 4.51 (t, 1H);4.84 (t, 1H); 5.27 (m, 2H); 5.96 (m, 2H).

Intermediate 287:(2S,5R)-6-(allyloxy)-3-(2-methoxyethyl)-2-(methoxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one

To a solution of(2S,5R)-6-(allyloxy)-3-(2-hydroxyethyl)-2-(hydroxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 286, 0.446 g, 1.75 mmol) and iodomethane (0.655 mL, 10.52mmol) in DMF (8 mL) at 0° C. was added sodium hydride (60% in mineraloil) (0.210 g, 5.26 mmol). The reaction mixture was stirred for 10minutes at 0° C., then quenched with water and diluted with ethylacetate. The layers were separated and the aqueous was extracted oncewith DCM. The combined organics were dried over magnesium sulfate,filtered and concentrated. Silica gel chromatography (0%-50% ethylacetate/hexanes) afforded the title compound (0.138 g, 28%) as a yellowoil.

MS: 283 ES+ (C₁₄H₂₂N₂O₄)

¹H NMR (300 MHz, DMSO-d₆) δ 2.12 (m, 2H), 3.02 (m, 1H), 3.21 (s, 3H),3.27 (s, 3H); 3.35 (m, 3H); 3.66 (m, 3H); 3.90 (m, 1H); 4.33 (m, 2H);5.27 (m, 2H); 5.96 (m, 2H).

Intermediate 288: (E)-triphenyl(prop-1-en-1-yl)phosphonium(2S,5R)-3-(2-methoxyethyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-3-(2-methoxyethyl)-2-(methoxymethyl)-1,6-diazabicyclo[3.2.1]oct-3-en-7-one(Intermediate 287, 0.177 g, 0.63 mmol) according to the proceduredescribed for Intermediate 17. The desired product was obtained as alight yellow foam (0.328 g, 84%).

MS: 323, 303 ES+ (C₁₄H₂₂N₂O₄, C₂₁H₂₀P)

Example 29(2S,5R)-2-(((1,5-dihydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl)carbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

To a solution of(2S,5R)-2-(((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)carbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate (Intermediate 296, 58.6 mg, 0.08 mmol) in anisole (906μl, 8.34 mmol) at 0° C. was added trifluoroacetic acid (321 μl, 4.17mmol). The reaction mixture was allowed to warm to room temperature andstir for 2.5 hours. The reaction mixture was diluted with DCM and water.The layers were separated and the aqueous extracted once more with DCM.The aqueous layer was directly loaded onto a 5.5 g RediSep Gold C18column and washed through with 100% water. Fractions 1 and 2 containeddesired product and were combined and lyophilized to afford a lightorange solid. The compound was run through another C18 column with waterand collected in tubes 1 and 2. They were combined and lyophilized. HPLCpurification (YMC Carotenoid C30 21.2 mm×150 mm 5 μm coupled withSynergi Polar RP 100 mm×21.2 mm 4 μm, 0%-15% acetonitrile/water)afforded the title compound as a pink solid (3.4 mg, 9.3%) afterlyophilization.

MS: 417 ES+ (C₁₄H₁₆N₄O₉S)

¹H NMR (300 MHz, DMSO-d₆) δ: 1.93 (s, 3H); 3.31 (m, 1H); 3.61 (m, 1H);4.22 (m, 1H); 4.49 (m, 2H); 4.61 (m, 1H); 5.69 (m, 1H); 6.49 (s, 1H);7.64 (s, 1H).

The Intermediates for Example 29 were Prepared as Follows Intermediate289: 2-(hydroxymethyl)-5-[(4-methoxybenzyl)oxy]-4H-pyran-4-one

To a stirred slurry of kojic acid (500 g, 3.518 mol) in dry DMF (5 L)was added dry potassium carbonate (972 g, 7.036 mol) followed by4-methoxy benzyl chloride (661 g, 4.221 mol) at 0° C. The reactionmixture was warmed to room temperature and then heated at 80° C. for 3hours. After cooling to room temperature the reaction mixture was pouredinto ice cold water (15 L) and stirred vigorously. The precipitate wascollected by filtration and dried under vacuum to afford the titlecompound as a pale brown solid (687 g, 75%).

¹H NMR (300 MHz, DMSO-d₆) δ 3.74 (s, 3H); 4.27 (s, 2H); 4.83 (s, 2H);5.68 (bs, 1H); 6.29 (s, 1H); 6.93 (d, 2H); 7.33 (d, 2H); 8.12 (s, 1H).

Intermediate290:1-hydroxy-2-(hydroxymethyl)-5-[(4-methoxybenzyl)oxy]pyridin-4(1H)-one

To a stirred solution of2-(hydroxymethyl)-5-[(4-methoxybenzyl)oxy]-4H-pyran-4-one (Intermediate289, 101.7 g, 0.388 mol) in pyridine (1.35 L) was added hydroxylaminehydrochloride (134.7 g, 1.94 mol). The reaction mixture was heated at85° C. for 4 hours, then evaporated to dryness and triturated with water(700 mL). The precipitate was collected by filtration and washed withwater (250 mL). The solids were then stirred in isopropanol (100 mL) for12 hours, collected by filtration and dried under vacuum to afford thetitle compound as a white solid (41.2 g, 38%).

¹H NMR (300 MHz, DMSO-d₆) δ3.75 (s, 3H); 4.55 (s, 2H); 5.05 (s, 2H);6.95 (d, 2H); 7.06 (s, 1H); 7.38 (d, 2H); 7.47 (m, 1H); 8.25 (s, 1H);8.62 (d, 1H).

Intermediate 291:1-(diphenylmethoxy)-2-(hydroxymethyl)-5-[(4-methoxybenzyl)oxy]pyridin-4(1H)-one

To a stirred ice cold solution of1-hydroxy-2-(hydroxymethyl)-5-[(4-methoxybenzyl)oxy]pyridin-4(1H)-one(Intermediate 290, 45 g, 0.162 mol) was added potassium tert-butoxide(18.2 g, 0.162 mol), followed by benzyl chloride (36.14 g, 0.178 mol).The reaction mixture was stirred at room temperature for 12 hours, thenpoured into ice cold water (3 L) and stirred for 1 hour. The precipitatewas collected by filtration and dried under vacuum to afford the titlecompound as a pale brown solid (70.4 g, 98%).

¹H NMR (300 MHz, DMSO-d₆) δ 3.76 (s, 3H); 4.25 (d, 2H); 4.65 (s, 2H);5.54 (bs, 1H); 6.05 (s, 1H); 6.51 (s, 1H); 6.94 (d, 2H); 7.25 (d, 2H);7.41 (m, 11H).

Intermediate 292:2-((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)isoindoline-1,3-dione

To a solution of1-(benzhydryloxy)-2-(hydroxymethyl)-5-((4-methoxybenzyl)oxy)pyridin-4(1H)-one(Intermediate 291, 2 g, 4.51 mmol), phthalimide (0.664 g, 4.51 mmol) andtriphenylphosphine (1.178 g, 4.51 mmol) in THF (20 mL) at roomtemperature was added diisopropyl azodicarboxylate (2.397 mL, 12.18mmol). Reagents are insoluble. DMF (10 mL) was added. The reaction wasstirred at room temperature overnight. The reaction mixture was filteredand concentrated onto silica gel. Silica gel chromatography (0%-4%methanol/dichloromethane) did not offer separation of desired productfrom impurities. Fractions were combined and repurified. Silica gelchromatography (0%-30% acetone/dichloromethane) afforded the titlecompound (1.66 g, 64.3%) as a light yellow foam.

MS: 573 ES+ (C₃₅H₂₈N₂O₆)

¹H NMR (300 MHz, DMSO-d₆) δ 3.76 (s, 3H); 4.51 (s, 2H); 4.70 (s, 2H);5.71 (s, 1H); 6.63 (s, 1H); 6.94 (m, 2H); 7.26 (m, 2H); 7.45 (m, 10H);7.62 (s, 1H); 7.88 (m, 4H).

Intermediate 293:2-(aminomethyl)-1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)pyridin-4(1H)-one

To a solution of2-((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)isoindoline-1,3-dione(Intermediate 292, 1.66 g, 2.90 mmol) in chloroform (20 mL) and methanol(10 mL) at room temperature was added hydrazine monohydrate (0.284 mL,5.80 mmol). The reaction mixture was stirred overnight at roomtemperature. Another 1 eq of hydrazine monohydrate was added. After 3hours still see starting material. Added another 1 eq of hydrazinemonohydrate. After 2 hours the reaction mixture was filtered to removesolids. The filtrate was concentrated. The residue was triturated withMeOH and ether and the solids filtered off. This was repeated twicemore. The resulting solid was triturated once with chloroform and MeOHand the solids removed by filtration. The filtrate was concentrated toafford a yellow foam (1.01 g, 79%).

MS: 443 ES+ (C₂₇H₂₆N₂O₄)

¹H NMR (300 MHz, DMSO-d₆) δ 3.76 (s, 3H); 4.25 (d, 2H); 4.65 (s, 2H);5.54 (bs, 1H); 6.05 (s, 1H); 6.51 (s, 1H); 6.94 (d, 2H); 7.25 (d, 2H);7.29 (s, 1H); 7.41 (m, 11H).

Intermediate 294:(2S,5R)-6-(allyloxy)-N-((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide

To a solution of(2R,5R)-6-(allyloxy)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxylicacid (Intermediate 32, 0.54 g, 2.27 mmol) in DMF (10 mL) at roomtemperature was added2-(aminomethyl)-1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)pyridin-4(1H)-one(Intermediate 293, 1 g, 2.27 mmol),O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (1.724 g, 4.53 mmol) and N,N-diisopropylethylamine(1.579 mL, 9.07 mmol). After 15 minutes the reaction mixture was dilutedwith ethyl acetate and washed with saturated sodium bicarbonate, brine,and 1/1 brine/water twice. The organics were dried over magnesiumsulfate, filtered and concentrated. Silica gel chromatography (0%-70%acetone/dichloromethane) afforded the title compound (0.704 g, 47%) as alight orange foam.

MS: 663 ES+ (C₃₈H₃₈N₄O₇)

¹H NMR (300 MHz, DMSO-d₆) δ 1.80 (m, 3H); 3.18 (m, 2H); 3.76 (s, 3H);3.85 (m, 1H); 4.06 (m, 2H); 4.26 (m, 1H); 4.37 (m, 2H); 4.68 (s, 2H);5.33 (m, 3H); 5.82 (s, 1H); 5.95 (m, 1H); 6.52 (s, 1H); 6.94 (m, 2H);7.24 (m, 2H); 7.42 (m, 10H); 7.47 (s, 1H).

Intermediate 295: (E)-triphenyl(prop-1-en-1-yl)phosphonium(2S,5R)-2-(((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)carbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylSulfate

The title compound was prepared from(2S,5R)-6-(allyloxy)-N-((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-ene-2-carboxamide(Intermediate 294, 0.704 g, 1.06 mmol) according to the proceduredescribed for Intermediate 17. Silica gel chromatography (0%-5%methanol/dichloromethane) afforded the title compound (0.379 g, 35.5%)as a yellow foam.

MS: 703, 303 ES+ (C₃₅H₃₃N₄O₁₀S, C₂₁H₂₀P)

Intermediate 296:(2S,5R)-2-(((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)carbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylHydrogen Sulfate Sodium Salt

The Dowex(R) 50WX8-100, ion-exchange resin (35 g, 0.38 mmol) wasconditioned by stirring for 2 hours in 2N sodium hydroxide (80 mL, 0.38mmol). The resin was then loaded into column and washed with water untilthe pH was 7. It was then washed with (1/1) acetone/water (˜100 mL),followed by water (˜100 mL). (E)-triphenyl(prop-1-en-1-yl)phosphonium(2S,5R)-2-(((1-(benzhydryloxy)-5-((4-methoxybenzyl)oxy)-4-oxo-1,4-dihydropyridin-2-yl)methyl)carbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate (Intermediate 295, 0.379 g, 0.38 mmol) was taken up inacetonitrile (˜2 mL), loaded on the resin and washed through with water.The desired product mass was seen in tubes 1-2 (mixed fractions), 3-14(clean fractions). The two mixed fractions were run through the columnagain. Tubes 22-23 (mixed fractions) and tubes 24-31 (clean fractions)were lyophilized. The desired product was obtained as an off-white solid(113.4 mg, 43%).

MS: 703 ES+ (C₃₅H₃₄N₄O₁₀S)

¹H NMR (300 MHz, DMSO-d₆) δ 1.78 (m, 3H); 3.19 (m, 2H); 3.36 (s, 2H);3.75 (s, 3H); 4.00 (m, 2H); 4.23 (m, 1H); 4.67 (m, 2H); 5.40 (m, 1H);5.83 (s, 1H); 6.50 (s, 1H); 6.93 (m, 2H); 7.24 (m, 2H); 7.41 (m, 10H);7.45 (s, 1H).

Example 30 Biological Activity

Minimum Inhibitory Concentrations (MICs) were determined by the brothmicrodilution method in accordance with the Clinical and LaboratoryStandards Institute (CLSI) guidelines. Clinical Laboratory StandardsInstitute: Methods for Dilution Antimicrobial Susceptability Tests forBacteria That Grow Aerobically (8^(th) Ed. (2009)) M07-A8. In brief,organism suspensions were adjusted to a 0.5 McFarland standard andfurther diluted to yield a final inoculum between 3×10⁵ and 7×10⁵colony-forming units (CFU)/mL. Bacterial inocula were made in sterile,cation adjusted Mueller-Hinton Broth (Beckton Dickinson) for eitherEscherichia coli, Klebsiella pneumoniae, or Pseudomonas aeruginosa.Haemophilus influenzae bacterial inocula were made in sterile, cationadjusted Mueller-Hinton Broth (Beckton Dickinson) containing 0.5% yeastextract (Beckton Dickinson) plus 15 μg/ml Bovine Hematin (Sigma) and 15μg/ml β-nicotinamide adenine dinucleotide (Sigma). Bacterial inoculawere made in sterile, cation adjusted Mueller-Hinton Broth (BecktonDickinson) for Staphylococcus aureus or sterile, cation adjustedMueller-Hinton Broth containing 2.5% lysed horse blood (Hema Resource &Supply Inc.) for Streptococcus pneumoniae and Streptococcus pyogenes. Aninoculum volume of 100 uL was added to wells (using a Tecan EVO robot)containing 2 uL of DMSO containing 2-fold serial dilutions of drug. Aninoculum volume of 100 μL was added to wells (using a Tecan EVO robot)containing 2 μL of DMSO containing 2-fold serial dilutions of drug. Allinoculated microdilution trays were incubated in ambient air at 35° C.for 18-24 hours. Following incubation, the lowest concentration of thedrug that prevented visible growth as read at OD600 nm was recorded asthe MIC. Performance of the assay was monitored by the use of laboratoryquality-control strains and commercially available control compoundswith defined MIC spectrums, in accordance with CLSI guidelines.

MIC (μg/mL) S. aureus S. aureus Ex. # S. pneumon. S. pyogenes (MSSA)(MRSA) H. influenzae E. coli K. pneumon. P. aeruginosa 1 6.25 3.1325 >200 ND 50 50 200 2 25 3.13 100 100 12.5 100 100 >200 14 100 100100 >200 12.5 >200 >200 >200 3 50 50 >200 >200 3.13 >200 >200 >2007 >200 >200 >200 >200 >200 >200 >20 >200 4 200 50ND >200 >200 >200 >200 >200 5 100 50 >200 >200 100 >200 >200 >200 12 2525 ND >200 100 100 50 >200 13 50 100 >200 >200 200 >200 >200 >200 100.39 <0.2 0.78 >200 0.78 12.5 12.5 200 6 25 25 200 >200 >200 >200 >20025 8 ND ND ND ND ND ND ND ND 9 ND ND ND ND ND ND ND ND 15 ND ND ND ND NDND ND ND 11 25 25 100 >200 >200 >200 >200 >200 16 50 50 100 >200 50 100100 >200

MIC (μg/mL) Pseudo- Klebsiella Pseudo- monas Pseudo- pneu- monasaeruginosa monas moniae aeruginosa PER-1, aeruginosa KPC2 AmpC OXA-10AmpC Ceftazidime 256 128 256 32 Ex. 13 >16 >16 >16 >16 Ex. 101 >16 >16 >16 Ex. 6 0.25 16 16 16 Ex. 4 >64 >64 >64 >64 Ex. 264 >64 >64 >64 Ex. 1 8 64 >64 >64 Ex. 11 >16 >16 >16 >16 Ex. 1264 >64 >64 >64 Ex. 8 >16 >16 >16 >16 Ex. 16 8 >16 >16 >16 Ex.9 >16 >16 >16 >16 Ex. 5 ND ND ND ND Ex. 15 ND ND ND ND Ex. 7 ND ND ND NDEx. 3 ND ND ND ND Ex. 14 ND ND ND ND

Example 31

Synergy with β-lactams was assessed against several organisms producinga variety of β-lactamases belonging to different classes. Bush & Jacoby,Updated Functional Classification of β-Lactamases, Antimicrobial Agentsand Chemotherapy, 54:969-76 (2010). In the assays, growth inhibitoryactivity (MIC) was measured using either two-fold dilutions of thepartner β-lactam at a fixed concentration of compounds of formula (I).Compounds were prepared in DMSO, and MIC determinations using thecompound of formula (I)/β-lactam combinations were done according to theCLSI guidelines. Clinical Laboratory Standards Institute: Methods forDilution Antimicrobial Susceptability Tests for Bacteria That GrowAerobically (8^(th) Ed. (2009)) M07-A8. Synergy was defined as afour-fold or more reduction in the MIC of the β-lactam in the presenceof the compound of formula (I), compared to the β-lactam alone.

MIC (μM) Pseudo- Klebsiella Pseudo- monas Pseudo- pneu- monas aeruginosamonas moniae aeruginosa PER-1, aeruginosa KPC2 AmpC OXA-10 AmpCCeftazidime 256 128 256 32 +Ex. 13 (4 ug/ml) ≦0.50 64 64 16 +Ex. 10 (4ug/ml) N 2 2 1 +Ex. 6 (4 ug/ml) N 4 2 2 +Ex. 4 (4 ug/ml) 2 64 64 32 +Ex.2 (4 ug/ml) 2 64 128 64 +Ex. 1 (4 ug/ml) ≦0.50 4 2 4 +Ex. 11 (4 ug/ml) 132 16 4 +Ex. 12 (4 ug/ml) ≦0.50 64 32 16 +Ex. 8 (4 ug/ml) ≦0.50 64 4 8+Ex. 16 (4 ug/ml) ≦0.50 32 32 16 +Ex. 9 (4 ug/ml) ≦0.50 64 16 32 +Ex. 5(4 ug/ml) ND ND ND ND +Ex. 15 (4 ug/ml) ND ND ND ND +Ex. 7 (4 ug/ml) NDND ND ND +Ex. 3 (4 ug/ml) ND ND ND ND +Ex. 14 (4 ug/ml) ND ND ND ND +Ex.17 (4 ug/ml) 16 ND ND ND +Ex. 18 (4 ug/ml) 16 ND ND ND +Ex. 19 (4ug/ml) >32 ND ND ND +Ex. 20 (4 ug/ml) 15 ND ND ND +Ex. 21 (4 ug/ml) 16ND ND ND +Ex. 22 (4 ug/ml) 4 ND ND ND +Ex. 23 (4 ug/ml) >256 ND ND ND+Ex. 24 (4 ug/ml) 8 ND ND ND +Ex. 25 (4 ug/ml) >8 ND ND ND +Ex. 26 (4ug/ml) 4 32 32 16 +Ex. 27 (4 ug/ml) 16 64 4 8 +Ex. 28 (4 ug/ml) ND ND NDND +Ex. 29 (4 ug/ml) ND ND ND ND

Pseudo- Klebsiella Pseudo- monas Pseudo- pneu- monas aeruginosa monasmoniae aeruginosa PER-1, aeruginosa KPC2 AmpC OXA-10 AmpCpiperacillin >512 256 64 256 +Avibactam (4 8 32 32 32 ug/ml) +Ex. 1 (4ug/ml) N 16 16 16 ceftazidime 256 128 256 32 +Avibactam (4 ≦0.50 8 8 4ug/ml) +Ex. 1 (4 ug/ml) ≦0.50 4 2 4 +Ex. 10 (4 ug/ml) ≦0.50 2 2 2ampicillin >512 >512 >512 >512 +Avibactam (4 16 512 512 >512 ug/ml) +Ex.1 (4 ug/ml) 128 512 512 >512 aztreonam >512 64 128 32 +Avibactam (4≦0.50 8 16 8 ug/ml) +Ex. 1 (4 ug/ml) 2 4 8 8 +Ex. 10 (4 ug/ml) ≦0.50 816 8 cefepime 512 32 32 16 +Avibactam (4 ≦0.50 4 8 8 ug/ml) +Ex. 1 (4ug/ml) ≦0.50 4 8 8 Ceftaroline >512 256 64 256 +Avibactam (4 ≦0.50 16 3232 ug/ml) +Ex. 1 (4 ug/ml) 1 8 32 16 ceftriaxone >512 >512 128 512+Avibactam (4 ≦0.50 128 16 64 ug/ml) +Ex. 1 (4 ug/ml) ≦0.50 32 16 32meropenem 64 ≦0.50 4 32 +Avibactam (4 ≦0.50 ≦0.50 4 16 ug/ml) +Ex. 1 (4ug/ml) ≦0.50 ≦0.50 4 8 +Ex. 10 (4 ug/ml) ≦0.50 ≦0.25 4 8

The invention claimed is:
 1. A compound according to formula (Ia):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is —CONR′R″,—CN, or an C₁-C₃ alkyl substituted with C₁-C₃ alkoxy, —OH, —CN, —NR′R″,or —CONR′R″; R² and R³ are independently selected from H, halo, —CN,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₁-C₆alkoxy, —CONR′R″, or C(O)₂R′; wherein the alkyl, alkenyl, cycloalkyl,and alkoxy represented by R² or R³ are independently and optionallysubstituted by one or more halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl,C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″, 5-7 memberedheterocycle, —C(O)NR′R″ or —NR′C(O)R″; and each R′ and R″ areindependently selected from hydrogen, C₁-C₆ alkyl, C₃-C₆ cycloalkyl,phenyl, 5 to 6 membered heterocyclyl or a 5 to 6 membered heteroaryl;wherein each alkyl, cycloalkyl, phenyl, heterocyclyl and heteroaryl isoptionally and independently substituted with one or more halo, —CN,—OH, C₁-C₃ alkyl, C₁-C₃ haloalkyl, C₃-C₆ cycloalkyl, C₁-C₃ alkoxy, C₁-C₃haloalkoxy, —C(O)(C₁-C₆ alkyl), —C(O)(C₁-C₆ alkoxy), —NH₂, —NH(C₁-C₃alkyl), —N(C₁-C₃ alkyl)₂, a 5-7 membered heterocyclyl or a 5-7 memberedheteroaryl; provided that R² and R³ are not both hydrogen; and when R¹is —C(O)NR′R″, then neither of R² or R³ is —C(O)NR′R″.
 2. The compoundof claim 1, according to formula (III):

or a pharmaceutically acceptable salt thereof.
 3. The compound of claim2, or a pharmaceutically acceptable salt thereof, wherein R² and R³ areindependently selected from the group consisting of H, C₁-C₃ alkyl,C₃-C₆ cycloalkyl, and —CONR′R″, wherein the alkyl and cycloalkylrepresented by R² and/or R³ are independently and optionally substitutedby one or more group selected from halo, —CN, —OH, C₁-C₃ alkyl, C₁-C₃haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″, a siderophore,—C(O)NR′R″ and —NR′C(O)R″.
 4. The compound of claim 2, according toformula (V):

or a pharmaceutically acceptable salt thereof.
 5. The compound of claim4, or a pharmaceutically acceptable salt thereof, wherein R² is methyl,ethyl, isopropyl, or cyclopropyl, wherein each R² is optionally andindependently substituted with one or more group selected from —OH andC₁-C₃ alkoxy.
 6. The compound of claim 5, or a pharmaceuticallyacceptable salt thereof, wherein R² is methyl.
 7. The compound of claim2, according to formula (IV):

or a pharmaceutically acceptable salt thereof.
 8. The compound of claim7, or a pharmaceutically acceptable salt thereof, wherein R³ is C₁-C₃alkyl, C₂-C₆ alkenyl, C₃-C₆ cycloalkyl, or —CONR′R″, each of which isoptionally and independently substituted with one or more substituentselected from the group consisting of halo, —CN, —OH, C₁-C₃ alkyl,cyclopropyl, C₁-C₃ haloalkyl, C₁-C₃ alkoxy, C₁-C₃ haloalkoxy, —NR′R″, asiderophore, —C(O)NR′R″ and —NR′C(O)R″; and each R′ and R″ isindependently selected from H and C₁-C₃ alkyl.
 9. The compound of claim7, or a pharmaceutically acceptable salt thereof, wherein R³ is methyl,ethyl, isopropyl, cyclopropyl, —CONH₂, —CONH(C₁-C₃ alkyl), or —CON(C₁-C₃alkyl)₂, each of which is optionally and independently substituted withone or more group selected from —OH, C₁-C₃ alkyl, C₁-C₃ alkoxy, —NR′R″,C(O)NR′R″ and —NR′C(O)R″; and each R′ and R″ is independently selectedfrom H and C₁-C₃ alkyl.
 10. The compound of claim 7, or pharmaceuticallyacceptable salt thereof, wherein R³ is C₁-C₃ alkyl, cyclopropyl,—CONR′R″, wherein each alkyl, and cyclopropyl is optionally andindependently substituted with one or more —OH, C₁-C₃ alkoxy, —NH₂, or—NHC(O)(C₁-C₃ alkyl); and each R′ and R″ are independently selected fromH, C₁-C₃ alkyl, and 5-6 membered heterocyclyl, wherein each alkyl andheterocyclyl represented by R′ or R″ is optionally and independentlysubstituted with one or more —OH, C₁-C₃ alkyl, or C₁-C₃ alkoxy.
 11. Thecompound of claim 7, or pharmaceutically acceptable salt thereof,wherein R³ is methyl, —CH₂OCH₃, or —CONH₂.
 12. The compound of claim 2,or a pharmaceutically acceptable salt thereof, wherein R¹ is —CONR′R″,—CN, or an C₁-C₃ alkyl substituted with C₁-C₃ alkoxy or —OH; and the R′and R″ of R¹ are independently selected from the group consisting of H,C₁-C₃ alkyl, or a 5-7 membered heterocyclyl, wherein each alkyl andheterocyclyl of R′ and R″ is optionally and independently substitutedwith one or more —OH, C₁-C₃ alkyl, C₁-C₃ alkoxy, —NH₂, —NH(C₁-C₃ alkyl),—N(C₁-C₃ alkyl)₂, or a 5-7 membered heterocyclyl.
 13. The compound ofclaim 2, or pharmaceutically acceptable salt thereof, wherein R¹ is—CH₂OCH₃, —CONH(CH₂)-siderophore, —CONH₂,

and

represents the point of attachment to the bridged bicyclic core.
 14. Thecompound of claim 2, or a pharmaceutically acceptable salt thereof,wherein R¹ is —CH₂OCH₃ or —CONH₂.
 15. The compound of claim 2, or apharmaceutically acceptable salt thereof, wherein: R¹ is —CH₂OCH₃;—CONH₂, or

R² is —H or —CH₃; and R³ is —H, —CH₃, or —CONH₂; provided that R² and R³are not both H; and when R¹ is —CONH₂, or

then R³ is not —CONH₂.
 16. The compound:

or a pharmaceutically acceptable salt thereof.
 17. The compound:

or a pharmaceutically acceptable salt thereof.
 18. A compound of claim2, selected from the group consisting of:(2S,5R)-2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-cyano-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-4-methyl-7-oxo-2-(piperidinium-4-ylcarbamoyl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate;(2S,5R)-2-carbamoyl-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-cyano-4-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-(2-aminoethylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate;(2S,5R)-2-(methoxymethyl)-7-oxo-4-(prop-1-en-2-yl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-((5-hydroxy-4-oxo-1,4-dihydropyridin-2-yl)methylcarbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-carbamoyl-4-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsodium sulfate;(2S,5R)-2-carbamoyl-3-isopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsodium sulfate;(2S,5R)-4-carbamoyl-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate, monosodium salt;(2S,5R)-2,4-bis(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate Sodium salt;(2S,5R)-2-(1-(tert-butoxycarbonyl)piperidin-4-ylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate sodium salt;(2S,5R)-4-(dimethylcarbamoyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate sodium salt;(2S,5R)-2-(hydroxymethyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate Sodium Salt;(2S,5R)-3-methyl-7-oxo-2-(piperidin-1-ium-4-ylcarbamoyl)-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate;(2S,5R)-2-carbamoyl-3-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-4-(2-amino-2-oxoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-4-carbamoyl-2-(hydroxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium Salt;(2S,5R)-2-carbamoyl-3,4-dimethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-carbamoyl-3-ethyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-4-(2-aminoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate;(2S,5R)-2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-4-(2-acetamidoethyl)-2-carbamoyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylsulfate sodium salt;(2S,5R)-2-(methoxymethyl)-4-(methylcarbamoyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-2-carbamoyl-4-cyclopropyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt;(2S,5R)-3-(2-methoxyethyl)-2-(methoxymethyl)-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt; and(2S,5R)-2-(((1,5-dihydroxy-4-oxo-1,4-dihydropyridin-2-yl)methyl)carbamoyl)-4-methyl-7-oxo-1,6-diazabicyclo[3.2.1]oct-3-en-6-ylhydrogen sulfate sodium salt; or a pharmaceutically acceptable saltthereof.
 19. A pharmaceutical composition comprising a compound ofFormula (III), or a pharmaceutically acceptable salt thereof, as claimedin claim 2, and at least one pharmaceutically acceptable carrier,diluent, or excipient.
 20. The compound of claim 2, wherein R¹ is—CONR′R″ or —CN.